MtoZ Biolabs

Boston, United States

Protein analysis is a fundamental step in studying biological systems, and it is essential in various fields such as drug development, disease diagnosis, and agricultural breeding. At MtoZ Biolabs, we provide comprehensive protein analysis services using advanced mass spectrometry techniques combined with our expertise in proteomics. With the aim of advancing your research and maximizing your success, our protein analysis services are designed to be reliable, efficient, and precise.

Services at MtoZ Biolabs

Our protein analysis service is based on state-of-the-art mass spectrometry, including LC-MS/MS and MALDI-TOF/TOF. We utilize these technologies to identify and quantify proteins in biological samples. The proteins are first separated by HPLC and then ionized before being introduced into the mass spectrometer. The mass-to-charge ratios of the ions are measured to determine the molecular weight of the proteins. By comparing the obtained mass spectra to a database of known protein sequences, we can identify the proteins present in your sample.

Applications

Our protein analysis service can be applied in various fields, including but not limited to:

• Biomedical research: Identifying and quantifying proteins involved in diseases can help understand the disease mechanism and develop potential therapeutic targets.
• Drug development: Our service can assist in the discovery and validation of drug targets, as well as in the evaluation of drug efficacy.
• Agricultural breeding: Protein analysis can help to understand the genetic traits of crops and livestock, promoting the breeding of high-quality varieties.

Sample Submission Requirements

For successful protein analysis, please ensure your samples meet our submission requirements:

• Samples should be clear without visible impurities.
• The protein concentration should be higher than 1mg/mL.
• Ensure your samples are stored and shipped under appropriate conditions to prevent protein degradation.

At MtoZ Biolabs, we are committed to providing our clients with the most reliable and highest quality protein analysis service. Please feel free to contact us if you have any questions about our service.

About Us

MtoZ Biolabs is an integrate contract research organization (CRO) providing advanced proteomics, metabolomics, bioinformatics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. The name of MtoZ represents mass to charge ratio in mass spectrometry analysis, as most of our services are provided based on our well-established mass spectrometry platforms. Our services allow for the rapid and efficient development of research projects, including protein analysis, proteomics, and metabolomics programs.

MtoZ Biolabs is specialized in quantitative multiplexed proteomics and metabolomics applications through the establishment of state-of-the-art mass spectrometry platforms, coupled with high-performance liquid chromatography technology. We are committed to developing efficient, and effective tools for addressing core bioinformatics problems. With a continuing focus on quality, MtoZ Biolabs is well equipped to help you with your needs in proteomics, metabolomics, bioinformatics, and biopharmaceutical research. Our ultimate aim is to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample.

Email: marketing@mtoz-biolabs.com

In recent years, the use and number of biotherapeutics has increased significantly. For these protein-based therapies, the quantitation of aggregates is of particular concern, given their potential effect on efficacy and immunogenicity. This need has renewed interest in size-exclusion chromatography (SEC). SEC is a chromatographic method, which acts as a sieve and separates molecules in solution by their size, and in some cases molecular weight. Molecular sieves are materials containing precise and single tiny holes, which can be used to adsorb gases or liquids. SEC has long residence time for small molecular weight compounds into gel pores, whereas large molecular compounds are washed away earlier. The advantage of molecular sieve chromatography is that it can efficiently process at least 1nmol of target protein. The separation time is also very short, generally within 3 hours, and the chromatographic separation peak obtained at the same time is also smaller.

Advantages of SEC in Protein Purification

1. The buffer solution used for SEC can be exchanged and desalted.

2. Similar varieties (such as protein fragments and oligomers) can be separated.

3. Compatible with a variety of solvents.

4. Independent on any particular form of protein for preservation and elution.

The liquid chromatography is divided into positive and reverse phase chromatography according to the relative polarity of the mobile phase and the stationary phase. Reversed-phase liquid chromatography (RPLC) refers to chromatographic method that uses a hydrophobic stationary phase. RPLC has the characteristics of high resolution and high repeatability, therefore it has been widely used in protein and polypeptide analysis, esp. for the separation of small molecular weight proteins and protein fragments with molecular weight

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is a technology commonly used in protein purification and analysis. SDS-PAGE can separate proteins according to the differences in the charge and the different mobility due to different molecular sizes. If the protein sample has been highly purified and contains only one protein, the results would show a single protein band after SDS-PAGE separation. However, when there are multiple proteins in the protein samples, different proteins can be separated into multiple protein bands through SDS-PAGE. Therefore, SDS-PAGE technology provides a direct way to analyze the purity of protein samples. To meet the research needs of analyzing the purity of protein products, MtoZ Biolabs has developed and optimized a SDS-PAGE based workflow, and provides an accurate analytical service for protein/peptide purification study.

Analytical Principles of SDS-PAGE

SDS is a type of anionic surfactant that can break the hydrogen and hydrophobic bonds in proteins. SDS can bind to proteins in a certain proportion to form SDS-protein complexes, covering the intrinsic charges of proteins. Therefore, the migration speed of all kinds of SDS-protein complexes is only determined by the molecular weight of proteins. Different proteins are separated by SDS-PAGE electrophoresis, followed by protein staining and analysis of protein bands.

Reports

Experiment procedures

Parameters of SDS-PAGE

Protein purity results

Bioinformatics analysis

About Us

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

Email: marketing@mtoz-biolabs.com

MtoZ Biolabs uses LC-MS for antibody C-terminal variation analysis, including detection of the proportion of K deletion in the C-terminal end of antibody and other types of truncation on antibody C-terminal.

Glycosylation, especially N-glycosylation, is a universal post-translational modification in regarding the localization, function, activity of proteins in tissues and cells. Among the information related to N-glycosylation, the site of N- glycosylation is the basis for our understanding of its sugar chain function. Besides the application of point mutation in traditional biochemistry, the study of glycosylation sites has also played an important role in the identification of glycosylation sites in N-terminal. MALDI TOF MS and nano LC-ESI-MS/MS are commonly used to identify the N/O-glycosylation sites in the antibodies. MtoZ Biolabs uses both MALDI TOF MS and nano LC-ESI-MS/MS technology to provide efficient and accurate glycosylation analysis of multiple biopharmaceuticals, including proteins, antibodies, etc.

Resveratrol is a type of natural phenol, produced by several plants in response to injury, stresses, or attack by pathogens such as bacteria or fungi. The main function of resveratrol is to protect plant from invasion of pathogens. Resveratrol, as an important dietary supplements, has proven to help to extend life span and reduce cholesterol level. MtoZ Biolabs offers targeted resveratrol analysis service using an ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of resveratrol even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increasing detecting sensitivity.

List. Quantification of Resveratrol and Derivatives ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

About Us

MtoZ Biolabs is an integrate contract research organization (CRO) providing advanced proteomics, metabolomics, bioinformatics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. The name of MtoZ represents mass to charge ratio in mass spectrometry analysis, as most of our services are provided based on our well-established mass spectrometry platforms. Our services allow for the rapid and efficient development of research projects, including protein analysis, proteomics, and metabolomics programs.

MtoZ Biolabs is specialized in quantitative multiplexed proteomics and metabolomics applications through the establishment of state-of-the-art mass spectrometry platforms, coupled with high-performance liquid chromatography technology. We are committed to developing efficient, and effective tools for addressing core bioinformatics problems. With a continuing focus on quality, MtoZ Biolabs is well equipped to help you with your needs in proteomics, metabolomics, bioinformatics, and biopharmaceutical research. Our ultimate aim is to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample.

MtoZ Biolabs utilizes 2D-nano LC-MS/MS technology for protein identification. In our general workflow, protein complexes are separated in 1D/2D gel and the target protein is then digested into peptide fragments, followed by HPLC separation and tandem MS analysis. We use Scaffold and Mascot software for analyzing peptide MS data, and ensuring confident protein identification. MtoZ Biolabs provides integrate Protein Identification Service.

Experimental Instruments

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensityProtein Identifications and Intensity

Related Services

Protein Analysis

Protein Identification

Protein Mass Measurement

LC-MS Analysis of Pull-Down Proteins

Native MS Analysis

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein analysis is crucial for biochemical, and biopharmaceutical studies. To meet our customers needs in these fields, MtoZ Biolabs has developed a sophisticated mass spectrometry platform, and is proud to offer services covering identification of proteins, mass measurement and PTMs analysis of proteins, antibodies, and other biopharmaceuticals.MtoZ Biolabs provides integrate Protein Analysis Service.

Service Categories

Protein Identification

MtoZ Biolabs utilizes 2D-nano LC-MS/MS technology for protein identification. This service is compatible for analyzing both single and complex protein samples with high confidence.

Protein Mass Measurement

MtoZ Biolabs provides protein mass measurement service using MALDI-TOF technique. This service can be well applied for evaluation of sample purity, protein subunits and polymer complex.

Protein PTMs Identification

MtoZ Biolabs has established a powerful platform for analyzing multiple protein PTMs, including phosphorylation, glycosylation, acetylation, etc. This platform is highlighted by a wide detecting range and is suitable for detecting low abundance modified proteins.

LC-MS Analysis of Pull-Down Proteins

Native MS Analysis

Molecular mass is the most fundamental characteristics for proteins and peptides. It directly reflects the structural integrity of proteins, and hence it is commonly applied in identification of proteins and peptides. To better assist the scientific research of our clients, MtoZ Biolabs provides macromolecular mass measurement service based on Bruker ultrafleXtreme MALDI TOF/TOF, and high-resolution protein mass measurement service via nano LC-MS. This service can be applied to accurately analyze the molecular mass, subunits, and dimerization/polymerization of multiple biotechnological molecules, including proteins, peptides, antibodies, and vaccines, etc.MtoZ Biolabs provides integrate Protein Molecular Weight Determination Service.

Experimental Instruments

Applications

Measurement of the Molecular Mass of Protein/Peptide/Antibody/Vaccine, etc.Analysis of Number of Subunits in Protein SampleAnalysis of Dimerization/Polymerization of Protein SampleEvaluation of Sample Purity and Protein/Peptide Contaminants

Sample Submission Requirements

*Customers are recommended to contact us for detailed sample requirements before sending your samples.

Case Study

In this study, the molecular mass of the protein sample from one of our customer was analyzed by MALDI-TOF. The result below showed a very significant peak at 1046.6 Da with intensity of nearly 100%, which indicated the protein mass was 1046.6 Da with high purity. The measured protein mass was consistent with the theoretical value.

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensityProtein Identifications and Intensity

Related Services

Protein Analysis

Protein Identification

Protein Mass Measurement

LC-MS Analysis of Pull-Down Proteins

Native MS Analysis

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein primary sequence determines the advanced structure of a protein, influencing a proteins function. Therefore, it is essential to analyze the sequence of a protein, to verify the integrity and correct translation of a protein sample. MtoZ Biolabs has developed a comprehensive protein sequencing platform to meet various needs of our clients, from N-terminal and C-terminal sequencing to full-length and de novo sequencing. Our featured services are presented below, but if you have any special requirement, you are welcome to contact us for custom service.MtoZ Biolabs provides integrate Protein Sequence Analysis Service.

Service Categories

Protein De Novo Sequencing

De novo sequencing for analyzing sequences of novel protein, peptides, and antibodies, of which database information is not available.

Protein N-Terminal Sequencing

Accurate determination of N-terminal sequence of proteins and peptides, ensured by a combination of Edman Degradation Sequencing system and N-terminal De novo Sequencing technique.

Protein C-Terminal Sequencing

Comprehensive protein C-terminal integrity analysis, including sequence analysis, PTMs, and C-terminal truncation evaluation, based on high-resolution LC-MS/MS technology.

Protein Edman Degradation

Automatic Edman Degradation Sequencing system, equipped with HPLC technology, allowing analyzing over 30 amino acids at protein N-terminus.

Protein Full-Length Sequencing

Using LC-MS/MS technology and 6 digestive enzymes, MtoZ Biolabs provides true full-length sequencing service. Accurate sequence analysis and 100% of protein sequence coverage is guaranteed.

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

LC-MS Analysis of Pull-Down Proteins

Native MS Analysis

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein integrity is a crucial charateristics for protein, as protein truncation or microheterogeneity can lead to malfunction of protein. It is even more critical for the quality tests of biopharmaceutical products, such as antibodies and vaccines. Thus, analysis of the C-terminal sequence of protein products is necessary for evaluating the quality and function of protein products. MtoZ Biolabs have developed a LC-MS/MS system, which is suitable for analyzing both N-terminal sequence and C-terminal amino acid sequence and PTMs. We strictly follow the ICH Q6B Guidance to fulfill our customers needs.MtoZ Biolabs provides integrate C-Terminal Sequencing Service.

Analysis Workflow

Applications

Determination of N/C-Terminal Sequence of Proteins/Peptides/Antibodies/VaccinesAnalysis of PTMs

Sample Submission RequirementsDeliverables

Experimental ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Analysis Raw Data FilesN/C-Terminal Amino Acid Sequences

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein N-terminal is the start point of the expression of a protein. It also influences the subcellular distribution, degradation, and the turnover rate of a protein. Thus, sequence analysis of the N-terminal of a protein is very important for studying the function of a protein. Aimed to provide comprehensive sequencing service, MtoZ Biolabs has developed a sophisticated N-T sequencing platform consisting of Edman sequencing and protein N-T de novo sequencing, with their advantages complementary to each other.MtoZ Biolabs provides integrate N-Terminal Sequencing Service.

In this service, protein sequence is not restricted by sample types, N-terminal blockage, and PTMs. Protein N-T sequences can be accurately analyzed by Edman sequencing, whereas, the N-terminal blockage and PTMs can be analyzed by mass spectrometry.

Analysis Workflow

Experimental Instruments

Applications

Determination of N-Terminal Sequence of Proteins/Peptides/Antibodies/VaccinesAnalysis of PTMs

Sample Submission Requirements

Edman Sequencing

(1) Electroblotted Samples

Proteins separated by SDS-PAGE should be blotted onto PVDF membrane, nitrocellulose is not recommended. PVDF membrane can be stained with Coomassie blue or Poncaue red (Sliver stain is not recommended), followed by washing with ultra-pure water. The washing steps must be repeated several times, when glycine-buffer is used.

(2) Liquid Samples

1-10 ug amount of sample, with >90% purity. Avoid using Tris, glycine, guanidine, glycerol, sucrose, ethanolamine, SDS, Triton, X-100, Tween, and ammonium sulfate in the buffer.

MS Analysis

(1) Gel and liquid protein samples are acceptable.

(2) 5-10 ug protein samples are required. Sample purity should be as high as possible.

*Note

All reagents/solvent used must be of the highest purity to reduce contaminating substances. Samples should be handled with extreme caution and always in clean condition. Any source that may introduce contaminating proteins should be eliminated. Customers are welcome to contact us for detailed sample requirements before sending your samples.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensityProtein Identifications and Intensity

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Full-length sequence information is essential for studying the integrity and correct translation of biopharmaceuticals. In general, protein sequence analysis service provided by MtoZ Biolabs includes three steps: digestion of proteins into smaller peptide fragments; tandem LC-MS/MS analysis of peptides; MS/MS data analysis. Traditional protein sequencing only uses trypsin for digestion, which renders the final protein sequence coverage of around 60%. To ensure 100% protein sequence coverage, MtoZ Biolabs uses up to 6 kinds of protein enzymes for protein digestions, eliminating any missing peptides.MtoZ Biolabs provides integrate Protein Full-Length Sequencing Service.

Analysis Workflow

Determination of Full-Length Sequence of Proteins/Peptides/Antibodies/VaccinesAnalysis of PTMs & Chemical Modifications on ProteinVerification of the Correct Translation of Recombinant Proteins

Sample Submission Requirements

Case Study

In this project, we analyzed the full-length sequence of an antibody product sent from one of our customers. To ensure 100% sequence coverage, we selected 4 enzymes for protein digestion, based on the predicted antibody sequence. The sequence mapping results showed 100% sequence coverage.

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensityProtein Identifications and Intensity

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Accurate determination of protein sequences is fundamental in the development of commercial monoclonal antibodies, vaccines, and diagnostic kits. Traditional sequencing methods uses database searching for protein sequencing, and has limitations for sequencing the unknown proteins, which may not be included in the database. De novo sequencing is a novel sequencing methods, with no need for database searching, and therefore, is the optimal method for analyzing unknown proteins and antibodies. Based on the world's most advanced mass spectrometry instrument, Obitrap Fusion Lumos, and combined with rich experience in bioinformatics analysis, MtoZ Biolabs has established a whole new generation of de novo sequencing platform that can achieve accurate sequence analysis of monoclonal antibodies and proteins, independent of any database searching. The primary sequence of proteins and antibodies and any mutations on the sequence can be analyzed quickly and accurately, without any prior information of the protein.MtoZ Biolabs provides integrate De Novo Sequencing Service.

Advantage of De Novo Sequencing

Advanced High-Resolution Mass Spectrometry Technology100% Protein/Antibody Sequence Coverage95% Accuracy for Sequencing the First Amino AcidMinimum Amount of Sample Required

De novo Sequencing

Antibody De Novo Sequencing

MtoZ Biolabs is proud to offer antibody de novo sequencing service, using high-resolution Obitrap Fusion Lumos mass spectrometry.

Protein De Novo Sequencing

MtoZ Biolabs develops an advanced mass spectrometry platform and ensures high-efficient and accurate protein de novo sequence analysis.

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

LC-MS Analysis of Pull-Down Proteins

Native MS Analysis

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Monoclonal antibody plays a key role in the development of biopharmaceuticals and diagnostic kits. The primary structure of the antibody, especially the amino acid sequence of the CDR region, is the core of antibodys biological function. Accurate and rapid analysis of its complete sequence is of great importance for characterization of antibody products. Analysis of the primary antibody sequence is the basis for antibody development and humanization for developing antibody-based drugs. However, due to the variation and modification of antibody products, the sequence information of most antibodies is often not included in the available databases. De novo sequencing technology uses advanced data processing algorithm for sequence analysis, has a superior advantage versus database dependent traditional methods. Antibody sequence can be accurately analyzed using the de novo sequencing method, with no need for database searching.MtoZ Biolabs provides integrate De Novo Antibody Sequencing Service.

Based on the world's most advanced mass spectrometry instrument, Obitrap Fusion Lumos, and combined with rich experience in bioinformatics analysis, MtoZ Biolabs has established a new generation of antibody de novo sequencing platform to achieve accurate analysis of the primary antibody structure. This platform is suitable for sequence analysis of different subtype antibodies, such as IgG and IgM, and different types of antibodies, including fluorescently conjugated antibodies, immobilized antibodies, and antibodies from different species.

Analysis Workflow

Service Advantages

Full Antibody Sequence Coverage

To achieve 100% antibody sequence determination, MtoZ Biolabs selects 5 different proteases for enzyme digestion. 100% full sequence splicing of light and heavy chains of antibody molecules will be achieved by using complementarity between different cleaved peptide segments. The CDR sequence of the antibody light chain is required to be identified at least 10 times during sequence analysis in order to fully ensure the accuracy of the sequencing results (as shown in the figure below).

Advanced Data Processing Algorithm

Based on the literature published in Nature Biotechnology and professional proteomics magazine JPR, and combined with the professional PEAKS Mass Spectrometry Software, MtoZ Biolabs has developed our own mass spectrometry raw data processing method. This method enables accurate identification of antibody sequences without missing any valid data information.

High Precision and Accuracy

MtoZ Biolabs uses Orbitrap Fusion Lumos mass spectrometer with the highest resolution and sensitivity for antibody sequencing. The MS scanning speed of Orbitrap Fusion Lumos has been significantly improved for detecting more peptides, and increases the credibility of the results. Increased MS/MS scanning speed enables detection of more collision-induced fragments (shown below) and accurate determination of leucine and isoleucine can be achieved by analyzing the characteristic peaks in the MS/MS spectrum.

Figure 3. HCD Collision-Induced MS/MS Spectrum

Sequencing Multiple Types of Antibodies

Our well-established technology can effectively analyze antibody with modifications of small groups, such as FITC, Biotin, and Alexa. Although large protein modification groups may affect the accuracy of sequencing, the impact can be minimized if the concentration of protein sample is high enough.

Sequence Prediction and Strict Sequence Verification

We will first analyze the determined sequence to identify the CDR region of the antibody. At.

Protein sequences are critical information for study of protein function, as the amino acid sequence determines the advanced protein structures. Generally, there are two methods for protein sequence analysis, database search and de novo sequencing. Database search method identifies proteins by measuring peptide mass, and comparing to available protein databases. However, this method is unsuitable for analyzing antibodies, peptides, and proteins, of which the sequence information is either inaccessible, or not included in any databases. For these cases, de novo sequencing is the only way to decipher the protein sequence. MtoZ Biolabs uses the most advanced mass spectrometer-Orbitrap Fusion Lumos, for de novo sequencing service. Coupled with our professional bioinformatics analysis and reverse sequence confirmation, high-quality sequence analysis is guaranteed. Our de novo sequencing service is also well-suited for analyzing multiple kinds of antibodies, including IgM and IgG antibody subtypes, and antibodies modified with FITC, biotin, and Alexa, etc.MtoZ Biolabs provides integrate De Novo Protein Sequencing Service.

Analysis Workflow

Sequence Analysis of Protein/Peptide, of which Databases are UnavailablePTMs Analysis of Protein/Peptide, of which Databases are UnavailableSequence Analysis of CDR Region or Full-Length of Antibodies

Case Study

In this study, the full-length sequence of a monoclonal antibody is analyzed by our de novo sequencing system. MS/MS data in Fig. 1 indicated that complete fragmentation of peptide precursors and L/I residues can be easily discriminated after second collision. We used four enzymes (indicated by different color in Fig. 2) for protein digestion. Peptide mapping results showed complete sequence coverage with CDR being identified by at least 10 times, reaffirming accuracy of our sequence analysis.

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications, Intensity, and Peptide MappingProtein Sequence and PTMs Data

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho-Proteomics

Acetyl-Proteomics

Ubiquitin-Proteomics

Glyco-Proteomics

Disulfide Bond

Histone Modifications

Although hybridoma cells are immortalized, extensive passage of hybridoma cells may cause mutations on antibody genes, and hybridomas may loss their specificity to produce the correct antibody. Therefore, sequence information of antibody is of extreme importance for stable quality of antibody in large-scale industrial manufacturing. To solve this problem, MtoZ Biolabs has developed a PCR based antibody sequencing workflow, coupled with degenerate primer design. Compared with the mass spectrometry-based antibody sequencing method, this PCR-based antibody sequencing method is much more cost effective and less time consuming. In this method, degenerate FR1 primers are designed for PCR amplification of immunoglobulin (Ig) light and heavy chain variable regions from your hybridoma extracted mRNA. To better meet your research needs, we provide a full-package service, covering the whole analytical steps involving antibody sequencing. You can simply send us your hybridoma cells, and we will perform all the following analytical experiments.MtoZ Biolabs provides integrate PCR Based Antibody Sequencing Service.

Analysis Workflow

Sequencing of Antibodies Derived from Multiple Common Species, Including Human, Mice, Rat, etcQuick Turnaround TimeHigh Quality Analysis with Competitive PriceFull-Package Service and Custom Service Is Also Supported

Sample Submission Requirements

Deliverables

Amplified DNA Fragments of Light and Heavy Chain Variable Regions (Optional)Experimental Procedures and All ReagentsDNA Sequencing ResultsDNA Sequence Alignments of Heavy and Light Chain Variable RegionsDNA Gel ImagesAnalytical Raw Data Files

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho-Proteomics

Acetyl-Proteomics

Ubiquitin-Proteomics

Glyco-Proteomics

Disulfide Bond

Histone Modifications

Edman sequencing method analyzes the protein N-terminal sequence in sequential Edman reactions. While mass spectrometry technology has been commonly used for protein analysis, Edman sequencing is still a powerful and irreplaceable method for protein N-terminal sequencing. As a well-established sequencing method, Edman sequencing provides more accurate protein sequence data, compared to MS. MtoZ Biolabs is aimed at providing reliable protein analysis service. Our protein/peptide N-terminal sequence analysis is performed by SHIMADZU PPSQ-33A Analyzer, equipped with the newest HPLC analysis system.MtoZ Biolabs provides integrate Protein Sequencing Service by Edman Degradation.

Service Advantages

High Sensitivity in Detecting PTH-AA at Low Picomole LevelAnalyze N-Terminal 30 Amino AcidsNewest HPLC Analysis System, with UV_VIS SPD-20A DetectorAccurately Discriminate Amino Acids of High Similarity, i.e. I/L, and Q/K

Applications

Determination of N-Terminal Sequence of Proteins/Peptides/Antibodies/VaccinesVerification of the Correct Translation of Recombinant ProteinsVerification of the Sequence of Synthesized Peptides

Sample Submission Requirements

Electroblotted Samples: Proteins separated by SDS-PAGE should be blotted onto PVDF membrane. Nitrocellulose is not recommended. PVDF membrane can be stained with Coomassie blue or Poncaue red (Sliver stain is not recommended), followed by washing with ultra-pure water. The washing steps must be repeated several times, when glycine-buffer is used.Liquid Samples: 1-10 ug sample amount, with >90% purity. Avoid using Tris, glycine, guanidine, glycerol, sucrose, ethanolamine, SDS, Triton, X-100, Tween, and ammonium sulfate in the buffer.

*Note: All reagents/solvent used must be of the highest purity to reduce contaminating substances. Samples should be handled with extreme caution and always in clean condition. Any source that may introduce contaminating proteins should be eliminated. Customers are welcome to contact us for detailed sample requirements before sending your samples.

Deliverables

Experiment ProceduresParameters and Settings of Edman Degradation Sequencing SystemPTH-AA HPLC Raw Data FilesProtein N-Terminal Amino Acid Sequence

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho-Proteomics

Acetyl-Proteomics

Ubiquitin-Proteomics

Glyco-Proteomics

Disulfide Bond

Histone Modifications

In the life of an organism, the function of proteins is achieved by the interaction between proteins and proteins. Therefore, the study of new proteins can provide new directions for the study of the potential function of proteins. In many protein tests, pull-down assays are often used to test protein-protein-interaction. Pull-down's experiment uses highly purified and enriched bait protein to capture the targeted proteins that specifically interact with the bait protein in the cell, and greatly improve the efficiency of identification of new target protein. The specific steps of pull-down's experiment are to express the bait protein modified with poly-His, or Biotin tags. Using immobilized affinity ligand to link the bait protein to the solid-phase matrix, and then mix the bait protein with the protein sample. In this process, targeted protein specifically interacts with the bait protein, and thus, the targeted proteins are isolated from the protein sample mixture. This process can be used to isolate and concentrate a particular protein from a sample containing many thousands of different proteins. Identification of the target protein is then processed by the mass spectrometer. Using CST pull-down assay kit, combined with advanced UPLC-MS platform, MtoZ Biolabs provides quick and accurate pull-down protein analysis service.MtoZ Biolabs provides integrate Fusion Protein Interaction Analysis Service Pull-Down and MS Service.

1D SDS PAGE/2D PAGE Gel Samples

(1) Coomassie Blue, SYPRORuby and Silver staining can be all accepted for protein analysis.

(2) Since silver staining may not be compatible with the mass spectrometry analysis, we highly recommend you to use the following silver staining product:

ProteoSilver Plus, Sigma (Product # PROTSIL1 or PROTSIL2)

Dodeca Silver Stain, BioRad (Product # 161-0481 or 161-0480)

(3) No destaining treatment is needed for silver stained protein samples.

Liquid Samples

(1) There is no specific requirement for samples with amount of protein > 10 ug.

(2) For samples with amount of protein < 10 ug, SDS and salt should be reduced in the solution.

(3) Please provide information of the buffer system used for sample preparation and estimated amount of protein in the solution.

*Note: All reagents/solvent used must be of the highest purity to reduce contaminating substances. Samples should be handled with extreme caution and always in clean condition. Any source that may introduce contaminating proteins should be eliminated. Customers are welcome to contact us for detailed sample requirements before sending your samples.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensityProtein Identifications and Intensity

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho-Proteomics

Acetyl-Proteomics

Ubiquitin-Proteomics

Glyco-Proteomics

Disulfide Bond

Histone Modifications

Proteomics is a large-scale study of proteins in a certain organism. Depending on specific research objective, there are many analytical techniques for proteomics study. To better meet our clients needs, MtoZ Biolabs is proud to offer 5 quantitative proteomics service, including Label-free, iTRAQ/TMT, SILAC, MRM, and SWATH. We also provide custom proteomics service for specific project. You are welcome to contact us for more detailed information on project design and sample requirements.MtoZ Biolabs provides integrate Quantitative Proteomics Service.

Service Category

Label-Free

MtoZ Biolabs provides label-free proteomics service, featured by label-free property and scalable application for analyzing multiple samples at once.

iTRAQ/TMT

MtoZ Biolabs offers an all-inclusive iTRAQ/TMT proteomics service package, including in vitro isobaric labeling and LC-MS/MS analysis. This service is suitable for relative quantification analysis of up to 10 samples.

SILAC/Dimethyl

MtoZ Biolabs offers a complete set of SILAC/Dimethyl quantitative proteomics service, including in vivo metabolic labeling and LC-MS/MS analysis. This service is suitable for comparison of 2-3 samples.

SWATH

MtoZ Biolabs provides Sequential Window Acquisition of all Theoretical fragment ions (SWATH), featured by data independent acquisition with high-throughput quality and sensitivity comparable to that of MRM technique.

MRM

MtoZ Biolabs provides absolute targeted proteomics service, Multiple Reaction Monitoring (MRM) with the highest sensitivity and accuracy so far to meet various needs in high-quality quantitative proteomics study.

Related Services

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Label-free analysis is a powerful technique for identifying and quantifying relative changes in complex protein samples. In this method, samples are run individually and identified by MS/MS information. Relative quantitation of different samples is then analyzed by aligning chromatographic peak areas of precursor ions between various runs. The most distinctive advantage is that it quantifies proteins without any use of labels, and can analyze unlimited number of samples. Since Label-free is relatively simple and easy compared to other proteomics techniques, it has been widely used for proteomics study and biomarker discovery. MtoZ Biolabs is proud to offer Label-free quantitative proteomics service with faster simpler results. We can also perform MS3 analysis to ensure higher analytical accuracy of qualification and quantitation.MtoZ Biolabs provides integrate Label-Free Quantitative Proteomics Service, MS Based.

Analysis Workflow

Isotopic Labels are Not RequiredSimpler Experimental Procedures and Data AnalysisSamples are Run Individually, and Numbers of Samples are UnlimitedSuitable for Analyzing Various Types of Samples, Including Tissue, Cell, Blood, etc.

Sample Submission Requirements

*Note: For Co-IP protein samples, only 2-5 ug is required. During sample elution, detergents (such as SDS) and high oxygen concentration should be avoided. Alternatively, you can send the sample beads to us directly.

Bioinformatics Analysis

Label-Free MS Data Quality AssessmentMultivariate PCA AnalysisProtein Statistical Analysis: Venn Diagram, Volcano PlotFunctional Annotation: GO Annotation, KEGG Annotation, COG AnnotationClustering Analysis: Hierarchical Clustering, K-Means ClusteringNetwork Analysis: STRING Analysis

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and IntensitiesBioinformatics Analysis

Related Services

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho-Proteomics

Acetyl-Proteomics

Ubiquitin-Proteomics

Glyco-Proteomics

Disulfide Bond

Histone Modifications

Isobaric Tag for Relative Absolute Quantitation (iTRAQ) and Tandem Mass Tags (TMT) are two similar quantitative proteomic techniques developed by AB SCIEX and Thermo Fisher, respectively. The most distinctive advantage of these two techniques is that they can label up to 10 different samples and compare the relative proteomic change through LC-MS/MS analysis. Therefore, these techniques are widely applied to study proteomic changes under different treatments or in different development stages. MtoZ Biolabs is proud to offer iTRAQ/TMT analysis service to meet your research needs. On the basis of these two well-established techniques, we can also provide MultiNotch MS analysis service through MS3 or proton-transfer reactions (PTR) to further increase the accuracy of the proteomic quantitation. MtoZ Biolabs has developed a specialized platform, equipped with Q Exactive HF (Thermo Fisher), Orbitrap Fusion, Orbitrap Fusion Lumos mass spectrometers coupled with Nano-LC for iTRAQ/TMT analysis service. To better solve your research problems, we offer an all-inclusive service, including protein reduction and digestion, peptide labeling, LC-MS/MS analysis, MS data analysis and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your protein samples, and we will complete all the following experiments.MtoZ Biolabs provides integrate iTRAQ/TMT/MultiNotch Quantitative Proteomics Service.

The iTRAQ/TMT isobaric tags are similar and generally consist of three parts, reporter group, balance group, and reactive group. Peptides are labeled with isobaric tags by reacting with the reactive group. In MS spectrum, different iTRAQ-tag labeled peptides have the same mass, and thus appear in the same MS peaks. However, when fragmented in MS/MS, the reporter group dissociates to produce ion signals, which provide quantitative information regarding the relative amount of the peptide in the samples.

Analysis Workflow

High-Throughput: Analyze Up to 10 Samples at OnceIn Vitro Labeling: Compatible for Labeling Various Samples, Including Tissue, Cell, Blood, etc.High Sensitivity and Broad Detecting Range: Suitable for Analyzing Low Abundance ProteinsCompatible with Peptide Enrichment Experiment for PTMs Analysis

Applications

Analysis of large quantity of clinical samples, large scale of experimental analysis, and discovery of new biomarkers

Sample Submission Requirements

Bioinformatics Analysis

iTRAQ/TMT Data Quality AssessmentMultivariate PCA AnalysisProtein Statistical Analysis: Venn Diagram, Volcano PlotFunctional Annotation: GO Annotation, KEGG Annotation, COG AnnotationClustering Analysis: Hierarchical Clustering, K-Means ClusteringNetwork Analysis: STRING Analysis

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and IntensitiesBioinformatics Analysis

Related Services

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho-proteomics

Acetyl-proteomics

Ubiquitin-proteomics

Glyco-proteomics

Disulfide bond

Histone Modifications

Stable Isotope Labeling By Amino Acids In Cell Culture (SILAC) is a relative quantitative proteomic technique, which combines metabolic labeling with mass spectrometry analysis. In this technique, cell samples are labeled by culturing with light, middle, or heavy isotope labeled lysine and arginine. Since property of proteins is unaffected by the label, different cell samples are mixed at early experimental stage followed by LC-MS/MS analysis, eliminating any following intra-experimental variability involved by differential sample preparation process. Hence, SILAC is a powerful method to study the relative proteomic change under differential treatments.MtoZ Biolabs provides integrate SILAC/Dimethyl Quantitative Proteomics Service.

Similar to SILAC technique, Dimethyl labeling is also an isotopic labeling method. However, Dimethyl labeling is a chemical labeling process, and labels lysine or the N-terminal amino acid residues on peptide, instead of labeling proteins. Except for the difference in labeling method, SILAC and Dimethyl labeling technique have comparable sensitivity and accuracy, and are being more and more widely used in relative proteomics studies.

MtoZ Biolabs has developed a specialized platform, equipped with Q Exactive HF (Thermo Fisher), Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometers equipped with Nano-LC for SILAC/Dimethyl analysis service. To better solve your research problems, we offer an all-inclusive service, including cell culture, cell labeling, protein purification, digestion, peptide separation, LC-MS/MS analysis, and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your cell samples, and we will complete all the following experiments.

Analysis Workflow

Cell Samples: Frozen cells (shipped on dry ice), which can be passaged for at least 5 times.If you can finish the cell labeling and cell stimulation steps, we also accept labeled cell and protein samples. In this case, proteins should be of 50-100 ug/sample, shipped on dry ice.

Comparison of SILAC and Dimethyl labeling methods

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and IntensitiesBioinformatics Analysis

Related Services

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho-Proteomics

Acetyl-Proteomics

Ubiquitin-Proteomics

Glyco-Proteomics

Disulfide Bond

Histone Modifications

Sequential Window Acquisition of all Theoretical fragment ions (SWATH) is a brand new mass spectrometry technology co-developed by Dr. Ruedi Aebersold research group (ETH Zrich) and AB SCIEX. SWATH data is an independent acquisition (DIA) technique. Unlike data dependent acquisition technique, which only selects certain MS spectrum for MS/MS detection, SWATH analyzes all MS and MS/MS spectra, allowing detection of virtually all detectable proteins in a sample. In SWATH technology, MS spectra are divided into several narrow windows and peptides in each window are sequentially analyzed equally, followed by MS/MS analysis. SWATH technique combines the high-throughput quality of shotgun proteomics with high accuracy comparable to MRM. To better meet our clients needs, MtoZ Biolabs offers a comprehensive SWATH service, including optimized sample preparation, high-quality SWATH mass spectrometry analysis and professional bioinformatics analysis.MtoZ Biolabs provides integrate SWATH Based Protein Quantitative Service.

Analytical Platform

AB SCIEX Triple-TOF 5600-plus

Analysis Workflow

Quantitatively Analyze Nearly All Detectable Molecules, Including Low Abundance Proteins and PeptidesHigh Accuracy, Comparable to MRM LevelHigh-Throughput, Identify and Quantitate More than 2000 Proteins at OnceComplete Digital Archive of Sample in the First AnalysisPre-Experiment Method Development is Unnecessary

Sample Submission Requirements

Bioinformatics Analysis

SWATH Data Quality AssessmentMultivariate PCA AnalysisProtein Statistical Analysis: Venn Diagram, Volcano PlotFunctional Annotation: GO Annotation, KEGG Annotation, COG AnnotationClustering Analysis: Hierarchical Clustering, K-Means ClusteringNetwork Analysis: STRING Analysis

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and IntensitiesBioinformatics Analysis

Post-translational modifications (PTMs) are covalent and usually enzymatic modifications on proteins. Common PTMs include phosphorylation, acetylation, ubiquitination, glycosylation, and so on. These PTMs can cause huge influences on protein structure, distribution, and function, hence, increasing the complexity of proteome to a greater extent. Protein PTMs is generally analyzed by measuring the mass increased on the modified peptides. Since PTMs are generally present in very low abundance, specific enrichment procedures are required before PTMs identification. MtoZ Biolabs has established a powerful and professional PTMs analysis platform, which includes Thermo Fisher Q ExactiveHF and Obitrap Fusion Lumos mass analyzer system, coupled with Nano-LC system. Our aim is to provide the most professional support for our clients research.MtoZ Biolabs provides integrate Post-Translational Modifications Proteomics Service.

Analysis Workflow

PTMs Analysis Service Categories

Phospho ProteomicsMtoZ Biolabs provides protein phosphorylation analysis service with highly efficient phospho-peptides enrichment processing and Nano LC-MS/MS analysis. Coupled with SILAC/iTRAQ labeling, this service can be applied to large-scale phospho-proteomics analysis.

Acetyl ProteomicsMtoZ Biolabs provides precise acetyl-proteomics analysis service using Nano LC-MS/MS. We also use CST acetylation-specific antibodies for acetyl-peptide enrichment, and 2-3 different enzymes for protein digestion to ensure full scan of acetyl-peptides.

Ubiquitin ProteomicsMtoZ Biolabs offers protein ubiquitination identification and quantification service using high-resolution mass spectrometry analysis. We also use highly specific ubiquitin antibody for enrichment of low abundance ubiquitin-peptide.

Glyco ProteomicsMtoZ Biolabs utilizes the HCD/ETD mode of Orbitrap Fusion mass spectrometry for glycoprotein analysis. Coupled with Byonic software, we can accurately analyze N- and O-linked glycosylation sites and the corresponding glycogens.

Disulfide Bond AnalysisMtoZ Biolabs provides disulfide bond analysis service at both single protein level and proteome level. We have optimized our sample preparation method to reduce the chance of in vitro disulfide bond exchange and maintain the native protein structure to the greatest extent.

Histone Modification AnalysisMtoZ Biolabs has optimized our sample preparation protocols to obtain highly purified histones with the least effect to the modification, and ensures precise histone modification analysis using Nano LC-MS/MS.

*Note: We also provide custom analytical service to identify many other types of PTMs. For special requirement, please contact us for project discussion.

Sample Submission Requirements

*Customers are welcome to contact us for detailed sample requirements before sending your samples.

Case Study

In this study, the ubiquitination modification sites of a Co-IP protein sample are analyzed. Part of the final analytical results is listed as below, showing the peptide sequences and identified ubiquitination sites respectively.

Deliverables

Experiment Procedures 2. Parameters of Liquid Chromatography and Mass Spectrometer 3. MS Raw Data Files 4. Identification of PTM Modified Peptides

About Us

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

Email: marketing@mtoz-biolabs.com

Phosphorylation is one of the most important PTMs in cellular activities. It is a reversible process, regulated by various kinases and phosphatases respectively. Important as it is, phosphorylation takes part in regulating nearly all the cellular activities, including cell proliferation, differentiation, apoptosis, etc.MtoZ Biolabs provides integrate Quantitative Phosphoproteomics Service.

As most phosphoproteins are present in low abundance, and have wide dynamic range, enrichment procedures before mass spectrometry analysis are usually necessary for detecting phorsphoproteins. MtoZ Biolabs provides various enrichment methods, according to specific experimental objectives. TiO2 and IMAC enrichment can be employed for enriching the total phosphorylation proteome, and phosphor-Ser/Thr or phospho-Tyrosine antibodies can be applied to analyze specific targeted phosphoproteome. Additionally, we also provide iTRAQ/TMT protein labeling service to further increase the accuracy of quantitative phosphoproteomics analysis.

Based on our well-established phosphoproteome analysis platform, MtoZ Biolabs can provide an accurate all-inclusive service package, including protein purification, digestion, protein labeling, phosphoprotein enrichment, MS analysis, and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your cell samples, and we will complete all the following experiments.

Analysis Workflow

Service Advantages

High Accuracy: Phosphoproteome Enrichment and Sensitive MS AnalysisLess Intra-Sample Difference by Coupling With iTRAQ/TMT LabelingDetecting Low Abundance PhosphoproteinsDetecting Wide Dynamic Range

Sample Submission Requirements

*Note: We will perform testing experiments before we start official experiments. To ensure the most cost-effective and accurate analysis is provided, only qualified samples will proceed to the official analysis.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Sequence and Phospho-Site MappingBioinformatics Analysis

Related Services

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Ubiquitination is an important PTM in cell biology, as the ubiquitin-proteasome complex has mediated degradation of 80-85 percent proteins, and directly affects the expression level and function of proteins. MtoZ Biolabs is glad to offer ubiquitination identification and quantification service through high-resolution mass spectrometry analysis. For some low abundance ubiquitin-proteins, we can provide peptide enrichment service before mass spectrometry analysis. To ensure high specificity and efficacy of enrichment of ubiquitin-peptides, we use CST Ubiquitin antibody for enrichment process, and use 2-3 different enzymes for protein digestion to ensure full scan of ubiquitin-peptides.MtoZ Biolabs provides integrate Quantitative Ubiquitinomics Service.

MtoZ Biolabs has developed a specialized platform equipped with Q Exactive HF (Thermo Fisher), Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometers, equipped with Nano-LC for ubiquitin-proteomics analysis service. To better solve your research problems, we offer an all-inclusive service, including protein purification, digestion, peptide enrichment and separation, LC-MS/MS analysis, MS data analysis and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your cell/protein samples, and we will complete all the following experiments.

Analysis Workflow

Service Advantages

High-Throughput Analysis, Identify and Quantify Up Thousands of Proteins at Once 2. Quantitatively Analyze Low Abundance Ubiquitin-Proteins 3. Use 2-3 Enzymes for Protein Digestion, Ensure Full-Coverage Analysis 4. State-of-Art Nano LC and Orbitrap Fusion Mass Spectrometer

Sample Submission Requirements

*Note: We will perform testing experiments before we start official experiments. To ensure the most cost-effective and accurate analysis is provided, only qualified samples will proceed to the official analysis.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and Ubiquitin MappingBioinformatics Analysis

Multiple Reaction Monitoring (MRM) is a quantitative method for analyzing targeted proteomics. Based on the sequence information of targeted proteins, MRM selectively analyzes proteins correlating with the targeted peptide signals, eliminating signals from interfering peptides. There are three steps in MRM mass spectrometry analysis: isolation of targeted precursor peptides based on MS spectra, fragmentation of precursor peptides, and detection of fragmented ions based on MS/MS spectra.

MRM has the highest accuracy among all proteomics quantitation methods, enabling absolute quantitation of protein with reference to internal standard, and analysis of up to 200 proteins at the same time. MtoZ Biolabs is proud to offer an all-inclusive MRM service. Just tell us your project objective and send us the protein sample, we will perform all the corresponding experiments, and provide the most professional analytical service to meet your specific needs. MtoZ Biolabs provides integrate Multi Reaction Monitoring MRM Service.

Analytical Platform

AB SCIEX TripleTOF 5600, AB SCIEX Triple Quad 5500 and Q Exactive Fusion

Analysis Workflow

Service Advantages

Offers the Highest Specificity, Sensitivity, and Reproducibility for Proteomics Quantitation 2. Broad Dynamic Range, Covering 4 Orders of Magnitude of Protein Abundance 3. High-Throughput, and Can Identify Up to 200 Specific Proteins at Once 4. Absolute Quantitation of Multiple Proteins Without the Need of Antibody

Applications

Verification of Untargeted Proteomics Results, Such as Label-Free Proteomics 2. Absolute Quantitation of Up to 500 Proteins/Peptides 3. Analysis of Highly Homologous Protein Family Members 4. Protein Post-Translational Modification Analysis 5. Discovery and Absolute Quantitation of Biomarkers

Sample Submission Requirements

Bioinformatics Analysis

MRM Data Quality AssessmentMultivariate PCA AnalysisProtein Statistical Analysis: Venn Diagram, Volcano PlotFunctional Annotation: GO annotation, KEGG Annotation, COG AnnotationClustering Analysis: Hierarchical Clustering, K-means ClusteringNetwork Analysis: STRING Analysis

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and IntensitiesBioinformatics Analysis

Related Services

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Histones are highly conserved proteins, and histone subtypes have highly similar amino acid sequence. The N-terminal ends of histones can be modified by several types of PTMs, including acetylation, methylation, phosphorylation and so on, which affect the DNA transcription, replication, and DNA condensation, etc. Therefore, analysis on histone N-terminal modifications can provide valuable information for epigenetic studies. MtoZ Biolabs has optimized our sample preparation protocols to obtain highly purified histones with the least effect to the modification. We also use 2-3 enzymes for protein digestion to ensure full coverage analysis of histone sequence, eliminating any missing information of short peptides rendered by using only one enzyme.

MtoZ Biolabs has developed a specialized platform, equipped with Q Exactive HF (Thermo Fisher), Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometers, equipped with Nano-LC for histone modification analysis service. To better solve your research problems, we offer an all-inclusive service, including sample preparation, protein purification, digestion, peptide separation, LC-MS/MS analysis, histone modification analysis and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your cell samples, and we will complete all the following experiments. MtoZ Biolabs provides integrate Histone Post-Translational Modification Analysis Service.

Analysis Workflow

Sample Submission Requirements

*Note: We will perform testing experiment before we start official experiments. To ensure the most cost-effective and accurate analysis is provided, only qualified samples will proceed to the official analysis.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and PTMs MappingBioinformatics Analysis

Related Services

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Disulfide bond is a covalent bond formed between the sulfur atoms of two cysteines. This bond is crucial for the stability and function of proteins, and many secreted proteins or proteins that are located at the cellular surface have formed disulfide bond to maintain the stability. To meet specific research needs of our customers, MtoZ Biolabs provides disulfide bond analysis service at both single protein level and proteome level. We have optimized our sample preparation method to reduce the chance of in vitro disulfide bond exchange and maintain the native protein structure to the greatest extent.

MtoZ Biolabs has developed a specialized platform, equipped with Q Exactive HF (Thermo Fisher), Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometers, equipped with Nano-LC for disulfide bond analysis service. To better solve your research problems, we offer an all-inclusive service, including sample preparation, protein purification, digestion, peptide separation, LC-MS/MS analysis, pLink-SS software analysis and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your cell/protein samples, and we will complete all the following experiments. MtoZ Biolabs provides integrate Protein Disulfide Bonds Identification and Quantitative Analysis services.Analysis WorkflowApplications1. Mapping the Disulfide Bond Sites of Unknown Proteins2. Verify the Disulfide Bonds and Folding of Protein Samples

Service Advantages

Analysis of Proteins at Single Protein Level and Proteomics LevelHigh-Throughput, Quantitatively Identify and Analyze Disulfide Bridges of Multiple ProteinsProtein are Maintained and Analyzed in Native Status

Sample Submission Requirements

Note: Please use enough dry ice in the shipment, and choose express delivery to eliminate sample degradation. We will perform testing experiment before we start official experiments. To ensure the most cost-effective and accurate analysis is provided, only qualified samples will proceed to the official analysis.

Deliverables

Experiment Procedures2. Parameters of Liquid Chromatography and Mass Spectrometer 3. MS Raw Data Files4. Peptide Identifications and Intensities5. Protein Identifications and Disulfide Bonds Mapping6. Bioinformatics Analysis

Related Services

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Acetylation is a highly conserved and reversible post-translational modification. It mainly takes part in regulating gene expression through modifying nuclear histones, but can also regulate several metabolic enzymes and metabolism pathways. There has been over thousands of acetylated proteins identified in proteomics studies. However, most acetylated proteins has very low abundance, hence, enrichment of acetyl-peptides can help to increase the sensitivity of mass spectrometry analysis. We use CST acetylation-specific antibodies for acetyl-peptide enrichment, and use 2-3 different enzymes for protein digestion to ensure full scan of acetyl-peptides. Moreover, the MS analysis can also be coupled with iTRAQ/TMT labeling or SILAC labeling to further increase the accuracy of relative quantitation between samples.

MtoZ Biolabs has developed a specialized platform equipped with Q Exactive HF (Thermo Fisher), Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometers, equipped with Nano-LC for acetylproteomics quantitation service. To better solve your research problems, we offer an all-inclusive service, including protein purification, digestion, peptide labeling and enrichment, LC-MS/MS analysis, data analysis and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your samples, and we will complete all the following experiments. MtoZ Biolabs provides integrate Quantitative Acetylproteomics Service.

Analysis Workflow

Service Advantages

High-Through Analysis, and Quantitatively Analyze Several Sample at Once, When Coupled With Sample LabelingLarge Scale Analysis of Acetyl-Proteins, Including Low Abundance Proteins and PeptidesHighly Accurate Nano LC-MS/MS Detection SystemProfessional MS Data and Bioinformatics Analysis

Sample Submission Requirements

Note: We will perform testing experiment before official experiments begin. To ensure the most cost-effective and accurate analysis is provided, only qualified samples will proceed to the official analysis.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and Acetyl-Site MappingBioinformatics Analysis

Related Services

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Glycosylation is a post-translational modification, in which a carbohydrate is covalently added to protein. This modification serves a multiple crucial roles in cell biology, including cell recognition, signaling transduction, etc. However, glycosylation has a feature of high structural complexity, which increases the difficulty of analysis of glycoproteins. Although it has been predicted that about 50 percent proteins are glycosylated, so far, only 10 percent glycoproteins have been identified.

MtoZ Biolabs utilizes the HCD/ETD mode of Orbitrap Fusion mass spectrometry for glycoprotein analysis. Coupled with Byonic software, we can accurately analyze N- and O-linked glycosylation sites, and the corresponding glycogens. MtoZ Biolabs has also developed a specialized platform, equipped with Q Exactive HF (Thermo Fisher), Orbitrap Fusion, Orbitrap Fusion Lumos mass spectrometers, coupled with Nano-LC for glycoproteomics identification and quantification service.

To better solve your research problems, we offer an all-inclusive service, including protein purification, digestion, peptide labeling and enrichment, LC-MS/MS analysis, data analysis and bioinformatics analysis. All you need to do is to tell us your project objective, and send us your cell/protein samples, and we will complete all the following experiments. MtoZ Biolabs provides integrate Quantitative Glycoproteomics Service.

Analysis Workflow

Service Advantages

High-Throughput Analysis: Quantitatively Analyze Multiple Samples at Once, When Coupled With iTRAQ/TMT LabelingLarge Scale Analysis of Glycoproteins, Including Low Abundance Proteins and PeptidesState-of-Art Nano LC and Orbitrap Fusion Mass SpectrometerProfessional MS Data and Bioinformatics Analysis

Sample Submission Requirements

*Note: We will perform testing experiment before we start official experiments. To ensure the most cost-effective and accurate analysis is provided, only qualified samples will proceed to the official analysis.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Identifications and IntensitiesProtein Identifications and Glycogen MappingBioinformatics Analysis

Related Services

PTM Analysis

Phospho Proteomics

Acetyl Proteomics

Ubiquitin Proteomics

Glyco Proteomics

Disulfide Bond

Histone Modifications

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Protein Sequencing

Protein De Novo Seq

N-Terminal Sequencing

C-Terminal Sequencing

Edman Degradation

Protein Full-Length Sequencing

Microbial metabolomics is a study of the complete set of metabolites within a microorganism. Microorganism is a bunch of small creatures which can not be easily identified by naked eyes, and exist in multiple forms, including bacteria, fungi, and some algea, and have the most diverse metabolic characteristics. Each microorganism generally has relatively small genome and proteomes, and the genome and proteome data information are far less than that of the eukaryotes. However, the metabolome of each microorganism is very distinctive, and reflects the cellular activity in a more direct way. Thus, microbial metabolomics has been widely used in microbial taxonomy and function studies.MtoZ Biolabs provides integrate Microbial Metabolomics Service.

Based on Thermo Q Exactive and AB Q-TOF 5600 mass spectrometry systems, MtoZ Biolabs is proud to offer high-efficient and accurate microbial metabolomics analysis service. We design the most suitable sample preparation methods based on your requirements, and assure high-efficient metabolites extraction and highly accurate microbial metabolomics analysis.

Analysis Workflow

To obtain high-quality metabolomics analytical results, sample preparation method should fulfill the following requirements:

Metabolites Are Extracted from Microbial Sample and Structures Are Maintained to the Greatest ExtentExtraction Method Should Be Unbiased, and Does Not Change the Physical and Chemical Characteristics of Metabolites

*Note: Before we start official experiments, we will perform pre-experiments and test multiple extraction methods followed by mass spectrometry analysis. Based on the testing results, we make sure that the most efficient extraction method is selected for official experiments.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesMS Data Quality ChecksMetabolites Quantification DataBioinformatics Analysis (PCA, KEGG, etc.)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Targeted metabolomics is a study in which defined metabolites in a sample are identified and quantitatively analyzed. Quantitation and semi-quantitation of defined metabolites are undertaken through the use of internal standard compounds. Targeted metabolomics has an advantage of high specificity and accuracy. Thus, this method has been widely used to analyze and compare multiple targeted metabolites under different physiological states, and is a critical analytical method for discovery of new biomarker in metabolic diseases and study of early diagnosis of diseases.MtoZ Biolabs provides integrate Targeted Metabolomics Analysis Service.

MtoZ Biolabs offers targeted metabolomics analysis service using an LC-MS-based multiple reaction monitoring (MRM) and GC-MS-based single ion monitoring (SIM) metabolomics platforms, which have characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of targeted metabolites, even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increase the detecting sensitivity.

Analytical Steps of Targeted Metabolomics

Data Collection: Targeted Metabolites Standards Are Synthesized and the MRM Transition Values of the Standards Are Measured by Triple Quadrupole MSQuantitative Analysis of Samples Along with the Standards Using Triple Quadrupole MSStatistical Analysis and Bioinformatics Explanation: ANOVA, PCA, KEGG Annotation, etc.

Analysis Workflow

Sample Submission Requirements

Cells and Microbes Samples: Cellular activities should be terminated instantly, whereas maintaining the cell integrity.Animal Body Fluids Samples: Such as urine, blood, saliva, etc. Samples must be added with anticoagulant and preservative reagents right after sample collection, followed by freezing at -80C.Plant Tissues: 200 mg/sample. Samples should be frozen in liquid nitrogen right after sample collection, and then transferred to -80C for storage.Serum Samples: Repeated freezing and thawing of sample must be eliminated. Serum samples should be settled down at room temperature for 30 min in the collection tube, and then transferred to centrifuge tube and centrifuged at 8, 000 rpm, 5 min. After centrifugation, supernatant is aliquoted to freezing tubes with 500 uL/sample, and stored at -80C.Urine Samples: 1 mL/sample. Urine samples can be aliquoted to centrifuge tubes with 1 mL each tube, with addition 1/100 (w/v) sodium azide, and stored at -80C.Faeces Samples: 10 mg/sample.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesMS Data Quality ChecksMetabolites Quantification DataBioinformatics Analysis (PCA, KEGG, etc.)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

A carbohydrate is a biological molecule consisting of carbon, hydrogen and oxygen atoms, usually with hydrogen to oxygen atom ratio of 2:1. Carbohydrates are important components of organisms, and play multiple roles in regulating various biological activities, such as cell recognition and protein secretion. MtoZ Biolabs has established a platform for analyzing carbohydrates and metabolites in glycolysis pathway. We guarantee accurate analysis of metabolites in glycolysis pathways even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Carbohydrates Analysis Service.

List 1. Quantification of Low-MW Sugars

*All analytes are measured by UPLC-MRM/MS

List 2. Glycolysis / TCA / Nucleotide Analysis (Central Metabolism Profile)

*All analytes are measured by LC-ESI-QTOF mass spectrometer

List 3. Glycosylation/TCA/Nucleotide and NAD Related Metabolites Analysis

*All analytes and internal standards are measured by LC-ESI-QTOF mass spectrometer

List 4. TCA-Plus (Central Metabolism Profile)

Profile of Central Metabolism, including glycolysis, pentose-phosphate shunt, TCA cycle and nucleotide pools measured by LC-ESI-QTOF mass spectrometer.

List 5. Gly/TCA (Supplement to Central Metabolism Profile)

*All analytes and Internal Standards are measured by LC-ESI-QTOF mass spectrometer

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Carnitine is a quaternary ammonium compound involved in metabolism in most mammals, plants and some bacteria. Carnitine exists in two isoforms, D-carnitine and L-carnitine. Only L-carnitine exists in animals, and is involved in oxidation of fatty acids, therefore, L-carnitine has been added in diet pills to reduce body fat. MtoZ Biolabs uses ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex) for analyzing carnitine and derivatives. Our service enables highly accurate qualification and quantification analysis of over 355 kinds of carnitines. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Carnitines Analysis Service.

List 1. Quantification of Carnitines Using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

List 2. Quantification of Acylcarnitines Using ESI on a LC-QQQ

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Plant hormones, also named phytohormones, are chemicals that regulate plant growth. Opposite to animal hormones, plant hormones can be synthesized and secreted by every plant cell. Plant hormones not only regulate local cellular activities, and can also be transported to other part of the plant. Hormone receptors are highly sensitive in detecting hormones, therefore, hormones usually occur in extremely low concentration. Thus detection of hormones requires highly sensitive technology. MtoZ Biolabs has establish a platform consisting of NMR and ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which is able to detecting extremely low amount of plant hormones with high accuracy. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Plant Hormone Analysis Service.

List. Quantification of Plant Hormones

*All analytes and Internal Standards are measured by NMR and ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Bile acids are a type of steroid acid, which predominantly exist in the bile of animal and are nearly 80% of the organic compounds in the bile. There are two types of bile acids, primary and secondary bile acids. Primary bile acids are synthesized in the liver, and can be modified by taurine, glycine, sodium, and potassium salts to form multiple derivatives. Secondary bile acids are generated in the colon as a result of bacteria activities. The main function of bile acids is regulating digestion of dietary fats and oils and balance of energy. MtoZ Biolabs uses ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex) for bile acids analysis. Our service is compatible of analyzing multiple samples and is able for qualitatively and quantitatively analyzing over 90 kinds of bile acids.MtoZ Biolabs provides integrate Bile Acids Analysis Service.

List 1. Quantification of Unconjugated, Taurine- and Glycine- Conjugated Bile Acids by UPLC-MRM/MS

*All analytes and Internal Standards are measured by UPLC-MRM/MS

List 2. Quantitation of 50 Compounds in Bile Acids Analysis I (Above) and 16 Bile Acid Sulfates and Glucuronides in Plasma/Serum and Urine by UPLC-MRM/MS

List 3. Quantitation of 20 Human and Mouse Specific Bile Acids

List 4. Quantification of Bile Acids by Two-Step Solvent Extraction, Followed by RPLC Separation and Measurement by ESI- QQQ MRM

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Vitamins are fundamental substances constituting the human body, and essential for maintaining life activities. Although vitamins is vital for many biological activities, only limited amount of vitamins are needed. Vitamins have multiple functions in organisms. For example, vitamin D has a hormone-like function, and regulate mineral metabolism and tissue growth. Vitamin C and E function as antioxidants. Most vitamin B family members function as enzyme cofactors and regulate enzyme activities. MtoZ Biolabs has established a platform for analyzing multiple types of vitamins. With our well-established analytical platform and optimized sample preparation methods, we are able to accurately analyze over 70 types of vitamins even in low abundance.MtoZ Biolabs provides integrate Vitamins Analysis Service.

List 1. Vitamins Separated by RPLC and Analyzed by ESI-MS

List 2. Targeted Analysis of Water-Soluble Vitamins Using LC-MS

List 3. Targeted Analysis of Fat-Soluble Vitamins by LC-MS

List 4. Quantification of Endogenous Vitamin Metabolites Including Structural Isoforms and Vitamin-Like Compounds by UPLC-MRM/MS

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Hormone are a bunch of signaling molecules produced by glands in multicellular organisms that are transported by the circulatory system to target distant organs to regulate physiology and behavior. Hormones not only affect nearby cells and can also be transported to regulate distant cells. Moreover, hormones have many other functions. They take part in regulating a wide range of physiological processes, including respiration, digestion, development, and reproduction, etc. Therefore, hormones play systematical roles in life activities. MtoZ Biolabs uses GC/MS (7890A/5975C) and ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex) for animal hormones analysis. We guarantee accurate analysis of animal hormones even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Animal Hormones Analysis Service.

List. Quantification of Animal Hormones Using GC/MS (7890A/5975C) and ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Fatty acids are a type of carboxylic acids with a long aliphatic chain. Depending on whether the constituent fatty acids contain carbon-carbon double bonds, fatty acids are classified into two types, saturated and unsaturated. Fatty acids are considered as energy pool for organisms, as fatty acids produce large quantity of ATP when metabolized. Based on GC/MS (7890A/5975C) technology and coupled with fatty acid standards, MtoZ Biolabs can identify and quantitate over 40 kinds of fatty acids in your sample. We guarantee accurate analysis of fatty acids molecules even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Fatty Acids Analysis Service.

List. Quantification of Fatty Acids Using GC/MS (7890A/5975C)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Flax-lignans is a type of natural phytoestrogens, and has similar structure to human estrogen. This chemical mainly exists in flax seeds. According to previous studies, flax-lignans can help to prevent prostatic cancer and osteoporosis. MtoZ Biolabs offers targeted flax-lignans analysis service using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex). We guarantee accurate analysis of flax-lignans even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Lignans Analysis Service.

Quantification of Flax-Lignans Using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Polyphenols, also known as polyhydroxyphenols, are a structural class of mainly natural organic chemicals characterized by the presence of large multiples of phenol structural units. Polyphenols play an important role in the ecology of most plants, by regulating plant coloration, and also prevent microbial invasion. MtoZ Biolabs offers targeted polyphenols analysis service using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of polyphenols even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Polyphenols Analysis Service.

List 1. Identification & Quantification of Anthocyanins by HPLC-UV

List 2. Identification & Quantification of Polyphenols by LC-MS

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Flavones are a class of flavonoids based on the backbone of 2-phenylchromen-4-one. Flavones are mainly found in fruits, but also present in grape wines, and stem and herb seeds. Flavone plays an important role in life activities, thus it is also called vitamin P. MtoZ Biolabs offers targeted flavones analysis service using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has the characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of flavones even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Flavones Analysis Service.

List. Quantification of Flavone by ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Bioamine is a kind of low molecular weight compounds containing nitric fatty group or heterocycle group. Depending on the composition of amine, bioamine can be divided into two classes, monoamine and polyamine. Monoamine plays a role in muscular and blood vessel relaxation, and regulates cerebral cortex activity. Polyamine mainly promotes synthesis of macromolecules, including DNA, RNA, and proteins, and accelerates development and growth. MtoZ Biolabs offers targeted bioamine analysis service using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which have characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of bioamine even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Bioamine Analysis Service.

List. Quantification of Bioamine by ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Organic acid refers to organic compound with acidic properties. Organic acids mainly occur in the leaves, roots, and fruits of plants. Except for a few organic acids existing free states, most organic acids can bond with potassium, sodium, calcium, or certain organic acids to form organic salt. MtoZ Biolabs offers targeted organic acids analysis service using GC/MS (Agilent 7890A/5975C), which has the characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of organic acids even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Organic Acids Analysis Service.

List. Identification & Quantification of Organic Acids by GC-MS

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Aldehyde refers to organic compounds containing a functional group with the structure CHO. Traces of many aldehydes are found in essential oils and often contribute to their favorable odors. MtoZ Biolabs offers targeted aldehydes analysis service using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of aldehydes even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increase detecting sensitivity.MtoZ Biolabs provides integrate Aldehydes Analysis Service.

List. Quantification of Aldehydes by UPLC-MRM/MS

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Nucleotides are most well known as basic components and building blocks for nucleic acids, including DNA and RNA. Other than this, nucleotides also have various roles in biological activities. For instance, cAMP has been proven to be a secondary messenger and regulates downstream signaling pathways, and ATP helps to carry chemical energy to drive metabolism process. One of the derivatives of nuleotides, NAD, also takes part in energy metabolism through regulating electron transfer reaction. MtoZ Biolabs offers targeted nucleotides analysis service using GC/MS (7890A/5975C) and ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has characteristics of high accuracy, specificity, and sensitivity. Our service enables fast and accurate analysis of multiple nucleotides and derivatives, including ATP, cAMP and NAD, etc. We guarantee accurate analysis of nucleotides even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increased the detecting sensitivity.MtoZ Biolabs provides integrate Nucleotides Analysis Service.

List 1. Quantitation of NTPs and Cyclic-Di-AMP by UPLC-MRM/MS

List 2. Quantification of NAD and Derivatives by ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

MtoZ Biolabs: Analysis of Acetyl-CoA and Related Substances

Acetyl-CoA, an activated form of acetate, is formed by an acetyl group (CH3CO-) bonding to the thiol group of coenzyme A via a high-energy thioester bond. It is product of the decarboxylation of pyruvate produced by -oxidation and glycolysis of fatty acid (FA). In the initial step of the tricarboxylic acid cycle (TAC), the acetyl group is transferred to oxaloacetate to form citrate, and this cycle is also called the "citric acid cycle". MtoZ Biolabs utilizes ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex) for the quantitative and qualitative analysis of acetyl-CoA and its related substances.MtoZ Biolabs provides integrate Acetyl CoA Analysis Service.

Detection of Acetyl-CoA and Related Substances at MtoZ Biolabs

Our technicians, with extensive technical expertise, use ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex) alongside acetyl-CoA and related substance standards and isotopic standards for precise quantitative and qualitative analysis:

Figure 1. Services for Analysis of Acetyl-CoA and Related Substances

Sample Submission Requirements

Serum, plasma, urine, bile, bile acids; cell, animal tissues including liver, brain, and feces; plants, yeast, microorganisms, etc.

Sample requirements

(1) Blood, bile, etc.: 10 L

(2) Various tissues: 10 mg

(3) Feces, etc.: 10 mg

For other sample types and sizes, please contact MtoZ Biolabs salesman.

Sample Transportation

Considering the evaporation rate of 3-4 kg of dry ice per day, ensure ample dry ice used for mailing (preferably larger dry ice pieces as they evaporate slower), and sealed in foam boxes. Opt for the fastest possible mailing method to reduce sample degradation during transport.

Deliverables

Experimental ProceduresRelevant Mass Spectrometry ParametersQuantitative or Qualitative Information of Identified SubstancesMass Spectrometry ImagesRaw Data

Carotenoids are a type of organic pigments, existing in chloroplast, photosynthetic algea, and certain types of fungi and bacteria. As carotenoids can not be produced endogenously by animals, therefore, carotenoids must be incorporated through uptaking carotenoids-containing diet. MtoZ Biolabs offers carotenoids analysis service using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has the characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of carotenoids even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Carotenoids Analysis Service.

List. Quantification of Carotenoids by ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

A biomarker refers to a measurable indicator of some biological state or condition. Biomarkers are often discovered by evaluating and comparing metabolites under normal biological and pathogenic conditions, or pharmacologic responses to a therapeutic intervention. MtoZ Biolabs offers meat biomarkers analysis service using ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of meat biomarkers even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Carotenoids Analysis Service.

List. Quantification of Meat Biomarkers by ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

In organisms, except for a few existing in free states, most metals bind to biomolecules to form organic compounds or salts. Metals are extremely important for biological processes. Many metals act as cofactor for enzymes, and change the enzyme activities. MtoZ Biolabs offers metallomics analysis service using ICP-MS, which has characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of metals even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Metal/Metallomics Analysis Service.

List. Quantification of Metal/Metallomics by ICP-MS

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Amino acids are building blocks of proteins and essential components for organisms. Apart from this, amino acids are important for the nitrogen balance of the organism. Amino acids take part in nitrogen cycle through deamination processes, in which the amine group is removed from amino acids. MtoZ Biolabs offers amino acids analysis service using GC/MS (Agilent 7890A/5975C) and LC/MS/MS (AB Sciex 5500), which has characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of amino acid metabolites even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be greatly reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Amino Acid Analysis Service.

List. Quantification of Amino Acid Using GC/MS (7890A/5975C) and ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Tryptophan is an amino acid, which takes part in protein biogenesis. There are two sources of tryptophan, endogenous protein degradation and daily intake. Tryptophan plays vital role in biological activities, and has interaction with multiple biomolecules, including carbohydrates, proteins, vitamins, etc. MtoZ Biolabs offers tryptophan metabolites analysis service using an ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex), which has characteristics of high accuracy, specificity, and sensitivity. We guarantee accurate analysis of tryptophan metabolites even in low abundance. With our optimized sample preparation methods, interference from high-abundance dominant metabolites can be hugely reduced, thus further increasing detecting sensitivity.MtoZ Biolabs provides integrate Tryptophan Analysis Service.

List. Quantification of Tryptophan Metabolites by ACQUITY UPLC/TripleQuad5500 (Waters/AB Sciex)

Related Services

Metabolomics

Targeted Metabolomics

Untargeted Metabolomics

Lipidomics

Quantitative Proteomics

Label-Free

iTRAQ/TMT

SILAC/Dimethyl

SWATH

MRM

Protein Analysis

Protein Identification

Protein Mass Measurement

PTMs Identification

Lipidomics is a study of composition and quantification of lipids at a systematic level, so as to uncover the interactions between lipids and other macromolecules, and the mechanisms of lipids in regulating biological activities. At present, LC-MS and shotgun lipidomics are two mainstream technology for lipidomics analysis. LC-MS utilizes high-throughput liquid chromatography system to separate lipids based on their physical and chemical properties, followed by mass spectrometry analysis. Whereas, shotgun lipidomics is independent of liquid chromatography technology, and separates lipids through intrasource separation system, which adjusts pH value of solution and changes the ionization status of lipids, followed by direct infusion into mass spectrometer for lipids identification.MtoZ Biolabs provides integrate Lipidomics Analysis Service.

LC-MS is a powerful tool for discovery lipidomics, as LC-MS usually achieves detection of lipids at ppm level and accurate analysis of lipids side chains through MS2 fragmentation. Whereas, shotgun lipidomics is more widely used for (semi)-quantitation of targeted lipids. MtoZ Biolabs provides both LC-MS and shotgun lipidomics services to meet your specific needs.

Lipids Classes

Professional and Customized Experimental Design to Meet Your Research RequirementsMultiple Choices of State-of-the-Art Analytical Platforms for Reaching a Higher Level Both in Depth and ScopeLower Sample DemandStringent Quality Control System Covering the Entire Analytical Processes, Ensuring Acquisition of High-Quality DataCompatible with a Variety of Types of Samples, Ranging from All Kinds of Tissues and Cell Samples to Body Fluids, Etc.

Sample Submission Requirements

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesMS Data Quality ChecksMetabolites Quantification DataBioinformatics Analysis (PCA, KEGG, etc)

Drugs with biological origins are still the mainstream treatments for many diseases. These drugs include antibodies, recombinant proteins, vaccines, etc. With technological progression and expending demands, hundreds and thousands of new biopharmaceutical drugs have been developed each year. However, successful development of these drugs relies on ensuring the quality, potency, and safety of your biopharmaceutical product, as impurity and defection of these biopharmaceutical products may compromise the efficacy and place potential risks when applied in clinical stage. Therefore, evaluation of biopharmaceuticals is critical and necessary before clinical trials. To help solve the challenges in the developmental process of biopharmaceutical products, MtoZ Biolabs has established a comprehensive analytical platform for biopharmaceutical analysis, and is able to cover most biopharmaceutical related requirements. We strictly follow the International Conference on Harmonisation (ICH) guidelines of technical requirements for registration of pharmaceuticals for human use.MtoZ Biolabs provides integrate Biological Products Analysis Service.

Technical Platforms

Capillary Isoelectric Focusing

2D-DIGE

Size Exclusion Chromatography (SEC)

High Performance Liquid Chromatography (HPLC)

Reversed-Phase Chromatography (RP)

MALDI-TOF-MS/MS

ESI-TOF-MS/MS

Native MS

Hydrogen-Deuterium Exchange (HDX) MS

Identification of Biopharmaceuticals

Molecular Weight Analysis

N/C Terminal Sequencing

Peptide Mapping

Amino Acids Composition Analysis

Variation Analysis of Biopharmaceuticals

Glycan Profiling

Glycosylation Site Analysis

Disulfide Bond Analysis

Purity Analysis of Biopharmaceuticals

Purity AnalysisSDS-PAGE

Purity Analysis (SEC & RPLC)

Host Cell Protein (HCP) Analysis

Antibody-Drug Conjugate (ADC) Analysis

*Apart from the above services, we can also provide customized service for other types of biologics and biopharmaceuticals. You are welcome to contact us for specific requirement related to these biopharmaceuticals.

Service Advantages

Free Consultation and Experimental DesignWell-Established Analytical Platform & Comprehensive Range of TechnologiesFull-Package Service for Proteins/Antibody/VaccinesProfessional Data Analytical SystemSupport Custom Service

Applications

Antibody AnalysisAntibody-Drug-Conjugates (ADCs) AnalysisProtein/Peptide AnalysisVaccines AnalysisPEGylated Proteins AnalysisGlycan Analysis

At present, there are two types of mass spectrometry used for molecular weight analysis: Matrix-assisted laser desorption/ionization (MALDI-TOF) and Electrospray ionization (ESI-MS). The two methods have their own advantages and applications. MALDI-TOF can be used to directly measure the molecular weight of biopharmaceuticals, while ESI-MS analyzes multiple charge signals obtained by electric spray, and speculates the precise molecular weight through deconvolution. MALDI-TOF-MS is more suitable for analyzing biopharmaceuticals with molecular weight less than 25kDa. ESI-MS is recommended for molecular weight identification of protein substances with molecular weight of larger than 25kDa. MtoZ Biolabs has established a platform with both MALDI-TOF and ESI-MS technologies, which can be used to determine the molecular weight of all kinds of biopharmaceuticals, including antibodies, vaccines, proteins drugs, and polypeptides.MtoZ Biolabs provides integrate High-Resolution MS Molecular Weight Identification Service.

Figure 1. MALDI-TOF Analysis of Biopharmaceuticals

Figure 2. ESI-MS Analysis of Biopharmaceuticals

Sample Submission Requirements

Sample Requirement

The sample should be dissolved in the volatile solvent with a low content of salt and detergent.

Requirement for Concentration of DetergentsTransportation of Samples

Freeze-dried protein samples, transported at frozen or 4 .

Case Study

MALDI can analyze molecular weight of protein sample and the presence of polymers in protein or peptide segments in protein. The following is an actual result of determination of the molecular weight of a protein drug. We can see from the graph that there are dimers and trimmers existing in the protein sample.

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesMolecular Weight ResultsBioinformatics Analysis

Peptide mapping is a widely used approach for analyzing biopharmaceuticals, including antibody, protein and peptides. The common peptide mapping process only uses trypsin for protein digestion, and the average coverage rate of identified protein is about 60%. In order to obtain the full sequence information of the targeted protein, MtoZ Biolabs uses up to 6 kinds of proteases for protein digestion (Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C and Lys-N). After protein digestion, sequences of peptide are mapped for full-length sequence of biopharmaceuticals. MtoZ Biolabs has also developed LC-MS/MS based peptide mapping technology, ensuring accurate peptide mapping analysis. This technology can be used to analyze the sequence of biopharmaceuticals and confirm complete expression of recombinant protein. With well-established technology and professional expertise, we can guarantee 100% peptide coverage and accurate data analysis.MtoZ Biolabs provides integrate Peptide Mapping Service.

Figure 1. Peptide Mapping Analysis of Biopharmaceuticals

Analysis Workflow

Proper enzymes are selected for enzyme digestion and identification of target proteins.Peptide analysis through LC-MS/MS or UV LC-MS/MS.The total sequence of peptide coverage/peptide are mapped and compared with the theoretical sequence.Delivery of report.

Service Advantages

Combination of specific and nonspecific proteases, achieving coverage of 100% of any protein (polypeptide).Confirmation of each peptide is based on the MS/MS fingerprint of the sample, and is more accurate than MS analysis.Protein sample 100% sequence validation service.

Sample Submission Requirements

Gel and solution samples are all acceptable for peptide mapping service, but special pretreatment is required for gel sample.The required amount of protein sample: 10-20ug.Protein sample with purity of >80% is suggested.

Case Study

In the each and every experiment, we will try to improve identification efficiency and reduce the cost in accordance with the analytical principle. In this case, the customer sent us an in vitro expressed antibody for peptide mapping analysis and comparison with the theoretical sequence. After receiving the sample, we first selected 3 proteases for digestion and designed subsequent sequence analysis experiments based on the theoretical sequences provided by our customers. Part of the experimental data are shown below, which proved that the antibody obtained was consistent with the theoretical sequence.

In the final report, we will also provide detailed data on the coverage of the peptides as shown in the following graph.

Deliverables

Experiment ProcedureParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesPeptide Mapping ResultsBioinformatics Analysis

Amino acid composition analysis is a classical protein analysis method. It can be used to determine the composition and content of amino acids in protein/peptide, and non-typical amino acids in protein/peptide drugs. Analysis of amino acids is a necessary step before the complete sequencing of biopharmaceuticals, and provides valuable evidence for the analysis of the sequence of biopharmaceuticals. Based on amino acid derivative technology and high-efficiency liquid chromatography, MtoZ Biolabs provides efficient and accurate amino acid composition analysis service for biopharmaceutical products. MtoZ Biolabs provides integrate Amino Acids Composition Analysis Service.

The service mainly consists of two steps.

Hydrolysis of proteins and peptide samples into free amino acids.The derivation and detection of free amino acids.

MtoZ Biolabs uses phenyl isothiocyanate (PITC) for amino acid derivation, followed by a high-efficiency liquid chromatography for determining the composition and content of amino acid derivatives, so as to speculate the components of the amino acids in the protein/peptide samples.

Sample Submission Requirements

Protein sample should be dissolved in distilled water or PBS.Sample Quantity: We recommend you to provide 30ug total samples and aliquote the sample in 3 tubes.

Deliverables

Experiment ProceduresParameters of Liquid ChromatographyAmino Acid Composition Analytical ResultsBioinformatics Analysis

Related Services

Identification of Biopharm

Molecular Weight Analysis

N/C Terminal Sequencing

Peptide Mapping

Variation Analysis

Glycosylation Site Analysis

Disulfide Bond Analysis

Purity Analysis

Purity Analysis SDS-PAGE

Purity Analysis SEC and RPLC

Host Cell Protein (HCP) Analysis

Antibody-Drug Conjugate (ADC) Analysis

N- and C-terminal sequences are important structures and functional parts of protein and polypeptide, and they may play decisive roles in the biological function of protein. The main methods for protein C-terminal sequencing include carboxypeptidase, chemical and tandem mass spectrometry, and the common methods for N-terminal sequencing include Edman degradation sequencing and mass spectrometry. Each of these methods has the respective advantage and shortcoming. Therefore, the combination of a variety of different sequencing methods can adapt to the requirements of multiple protein sequencing. For example, Edman degradation may not well solve the problem of blockage and protein modification on the N-terminal end. This drawback can be overcomed by mass spectrometry analysis. MtoZ Biolabs uses both Edman degradation system and MALDI-TOF for N/C sequencing of biopharmaceutical products. This combination of two methods ensures accurate analysis of N-terminal blockage and modified protein N/C terminal. MtoZ Biolabs provides integrate N/C Terminal Sequencing Service.

Figure 1. Biopharmaceutical N/C-Terminal Sequencing

Sample Submission Requirements

Sample Types: Lyophilized & Solution SamplesAmount of Total Protein: >300 ugPurity: >90%Content of Salt: Volatile Salt

Many biotherapeutical products, including antibodies, vaccines, and recombinant proteins, are generated through biological processes. Despite of extensive purification steps employed, residual contaminants, as called host cell proteins (HCP), from the biological systems that are used to generate the biologics may still present in the final product. These HCPs, even of low levels, may compromise the safety and efficacy of the biotherapeutical products. Thus, detection and evaluation of HCP in biotherapeutical products are essential for quality and safety control.

MtoZ Biolabs uses 2D DIGE and Western blot technology for HCP analysis. This technology offers high sensitivity and can be employed for analyzing HCP from multiple species, such as mammalian cells, yeast, bacteria, and so on. Just tell us the potential HCP that may be contained in your sample, we will take charge of all the following analysis. MtoZ Biolabs provides integrate Host Cell Protein (HCP) Analysis Service.

Analysis Workflow

Service Advantages

Characterize HCP Originated from Multiple SpeciesComprehensive and Accurate Evaluation of Residual HCPWide Dynamic Range and High Sensitivity, Enabling Detection of Trace Amount of HCP ContaminantsHighly Efficient Analysis and Short Turnaround Time

Applications

Characterizations of HCP in Antibody ProductsDetection of HCP Contaminants in Recombinant ProteinsDetection of HCP Contaminants in Therapeutic Proteins and Vaccines

Deliverables

Information of Experimental Procedures and Reagents UsedParameters of 2D DIGE System and Mass SpectrometryHCP IdentificationRaw Data Files

Related Services

Identification of Biopharm

Molecular Weight Analysis

N/C Terminal Sequencing

Peptide Mapping

Amino Acids Composition

Variation Analysis

Glycosylation Site Analysis

Disulfide Bond Analysis

Purity Analysis

Purity Analysis SDS-PAGE

Purity Analysis SEC and RPLC

Antibody-Drug Conjugate (ADC) Analysis

Glycosylation, especially N-glycosylation, is a universal post-translational modification in regarding the localization, function, activity of proteins in tissues and cells. The study of the N-glycosylation site is an important prerequisite for understanding the function of sugar chain of the target protein. Among the information related to N-glycosylation, the site of N- glycosylation is the basis for our understanding of its sugar chain function. Besides the application of point mutation in traditional biochemistry, the study of glycosylation sites has also played an important role in the identification of glycosylation sites in N-terminal. MALDI TOF MS and nano LC-ESI-MS/MS are commonly used to identify the N/O-glycosylation sites in the antibodies. MtoZ Biolabs uses both MALDI TOF MS and nano LC-ESI-MS/MS technology to provide efficient and accurate glycosylation analysis of multiple biopharmaceuticals, including proteins, antibodies, etc. Upon the sample reception, Fc domain and Fab of the antibodies are first separated, followed by digestion into peptide fragments, and analysis of glycosylation site by MALDI TOF MS and nano LC-ESI-MS/MS. MtoZ Biolabs provides integrate Glycosylation Site Detection Service.

Figure 1. Glycosylation Site Analysis of Antibody

Analysis Workflow

Digestion of Biopharmaceutical Samples by Trypsin or Other Appropriate ProteasesSeparation of N-Glycopeptides and O-Glycopeptides Through Chromatographic ColumnDeglycosylation TreatmentPeptide Analysis by MALDI-TOF MS, or ESI-MS/MSAnalysis of the N/O-Glycosylation Site Based on the Mass Spectra ResultsComparison of the Analytical Results of Glycosylation Sites of Different Batch Samples

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesGlycosylation Site ResultsBioinformatics Analysis

Related Services

Identification of Biopharm

Molecular Weight Analysis

N/C Terminal Sequencing

Peptide Mapping

Amino Acids Composition

Variation Analysis

Disulfide Bond Analysis

Purity Analysis

Purity Analysis SDS-PAGE

Purity Analysis SEC and RPLC

Host Cell Protein (HCP) Analysis

Antibody-Drug Conjugate (ADC) Analysis

Disulfide bond (S-S bond) is formed by oxidation of the sulfhydryl group (-SH) on two cysteines in the protein. It is an important post-translational modification of protein. Disulfide bond is essential for protein molecules to maintain the correct advanced structure and maintain protein bioactivity. The distribution of disulfide bonds in antibody drugs is a direct structural characteristic of the drugs. Therefore, confirmation of disulfide bonds plays a very important role in the confirmation process of antibody drug structure. MtoZ Biolabs has developed a high-resolution mass spectrometry, coupled with pLink-SS software, to provide our customers with accurate analysis of disulfide bonds and free cysteines. Our sample preparation steps have also been optimized to prevent in vitro exchange of disulfide bonds, and maintain native structure. MtoZ Biolabs provides integrate Disulfide Bond Analysis Service.

Figure 1. Disulfide Bond Analysis of Biopharmaceuticals

Applications

Identification of the Number of Disulfide Bonds in the Biopharmaceuticals and Free CysteinesIdentification of the Position of Disulfide Bonds in the Biopharmaceuticals

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesDisulfide Bonds and Free Cysteines Analysis ResultsBioinformatics Analysis

Related Services

Identification of Biopharm

Molecular Weight Analysis

N/C Terminal Sequencing

Peptide Mapping

Amino Acids Composition

Variation Analysis

Glycosylation Site Analysis

Purity Analysis

Purity Analysis SDS-PAGE

Purity Analysis SEC and RPLC

Host Cell Protein (HCP) Analysis

Antibody-Drug Conjugate (ADC) Analysis

Antibody-Drug Conjugate (ADC) is a new type of biopharmaceutical product that is formed by linking a small molecular drug with potent cell toxicity, to monoclonal antibody. Antibody directs the ADC to target specific cancer cells and the small drugs function to kill the targeted cancer cells. Thus, the ADC has the characteristic of both the specificity of antibody and the toxicity of drugs to precisely remove cancer cells. Combining the advantage of both antibody and small drugs, ADC performs the best quality, reducing the potential damages to the greatest extent.MtoZ Biolabs provides integrate Antibody-Drug Conjugate (ADC) Analysis Service.

The preparation of ADC is commonly carried out through a two-step reaction.

Antibody reacts with a conjugate linker to generate an antibody-linker intermediate.The antibody-linker intermediate reacts with the small drug to form the final Antibody-Drug Conjugate.

The two-step reaction is shown in the following graph.

Problems That May Occur in the Antibody-Drug Conjugation Reaction

Due to unknown reasons, some of the antibodies and small molecules may not be successfully conjugated.Multiple numbers of binding sites in the antibody (Cys, Lys, etc.), may lead to heterogeneity of conjugation site and ratio of conjugation.Due to the hydrophobicity of small molecule drugs, different Drug antibody ratio (DAR) may lead to change of hydrophobicity of the final ADC.

As non-coupled small drugs are cytotoxic, these drugs along with free antibodies may affect the efficacy and safety of the ADC. Compared to monoclonal antibodies, the production process of ADCs drugs is more complex, therefore, in order to ensure the safety and effectiveness of the ADC, it is necessary to monitor the quality of the ADC. DAR is an important indicator to evaluate the production process and quality of products. Comprehensive evaluation of the ADC drug structure, DAR, efficacy and safety is critical before the ADC declaration. MtoZ Biolabs has developed an advanced platform for analyzing ADC, with a variety of high-resolution mass spectrometer analysis instruments. Combined with professional bioinformatics analysis team, we can quickly and accurately provide you with professional and systematic ADC analysis and evaluation services.

Analytical Platforms

MALDI-TOF-MS

ESI-TOF-MS

UV/VIS-MS

UV-MALDI-MS

RP-HPLC

HILIC

IEX

HIC

SEC

Antibody-Drug Conjugate (ADC) Analytical Programmes

MtoZ Biolabs provides one-stop systematical service to analyze ADC. The service contains three panels, including ADC characterization, potency analysis, and impurity analysis, ensuring systematic evaluation of the drug structure and efficacy of ADC.

Figure 2. Antibody-Drug Conjugate (ADC) Analytical Programmes

Deliverables

Experiment ProceduresParameters of Liquid Chromatography and Mass SpectrometerMS Raw Data FilesADC Evaluation ResultsBioinformatics Analysis

Monoclonal antibodies could recognize almost unlimited targets because their high specificity and affinity character. This ability has catapulted them as an attractive and fast-growing class of therapeutic & diagnostic agents over the past decades. Mapping the interactions between antibody (paratope) and antigen (epitope) is crucial for characterizing antibodys mechanism of action, predicting suitable antigens, as well as securing intellectual property. A number of different approaches have been used for antibody epitope mapping, including X-ray crystallography, NMR, mutagenesis, peptide-based ELISA, hydrogendeuterium exchange, etc. However, these methods generally require high levels of expertise and cost, or only give information on linear structures, which greatly impedes interpreting the precise nature of antibody-antigen interactions.MtoZ Biolabs provides integrate Antibody Epitope Mapping Service.

Services at MtoZ Biolabs

In order to meet this challenge, MtoZ Biolabs has introduced an innovative EpiLink technical platform. It is based on chemical cross-linking of proteins coupled with mass spectrometry analysis (CXMS) which can outline the overall protein architecture via identifying direct binding partners and localizing the binding interface. Yet, conventional CXMS may have problems detecting and analyzing interlinked peptides, the most informative category that faithfully reflects antigen-antibody interaction. Distinct from all current methods, our EpiLink Platform can offer multiple outstanding features.

Designed specifically for identifying and interpreting of cross-linking spectra, providing higher sensitivity and accuracy.Able to interpret conformational epitopes in original folding at amino acid level.Required very small sample volume.Fast and efficient, with a very short lead time frame.Professional team with rich experience in utilizing EpiLink antibody epitope analysisWith the help of EpiLink, we have successfully pinpointed the epitope information of multiple antibodies. Our state-of-the-art platform can be a robust tool to offer precise structural insights and support your research endeavors.

Related Services

Identification of Biopharm

N/C Terminal Sequencing

Peptide Mapping

Amino Acids Composition

Variation Analysis

Glycosylation Site Analysis

Disulfide Bond Analysis

Purity Analysis

Purity Analysis SDS-PAGE

Purity Analysis SEC and RPLC

Host Cell Protein (HCP) Analysis

Antibody-Drug Conjugate (ADC) Analysis

Protein mutation refers to the alteration in the amino acid composition or sequence of a protein, which subsequently leads to changes in the structure and function of the protein. Protein mutations can occur naturally or be induced artificially. The analysis of protein mutations primarily involves comparing the differences between normal proteins and mutant proteins, revealing the effects of mutations on protein structure, stability, function, and interactions with other molecules. In the fields of biotechnology and drug design, mutation analysis aids in understanding the molecular mechanisms of proteins, optimizing protein performance, predicting the impact of mutations on biological functions, providing a basis for biotechnology and drug design, and enhancing the production efficiency of bioproducts and the specificity of drug therapy.MtoZ Biolabs provides integrate Protein Mutation Analysis Service.

MtoZ Biolabs provides a one-stop, mass spectrometry-based protein mutation service utilizing Thermo's newly launched Obitrap Exploris 240 mass spectrometer combined with Nano-LC chromatography technology. Through high-resolution mass spectrometry analysis of protein amino acid sequence, it reveals whether the protein has been mutated or not, the site of mutation, and the amino acid sequences before and after the mutation, which can help research on the structure and function of proteins, the regulation of gene expression, and drug discovery.

Service Advantages

Advanced Mass Spectrometry Technology

We employ state-of-the-art high-resolution mass spectrometers, including the Q-Exactive series and Orbitrap series, to ensure the accuracy and reliability of data. These instruments capture extensive protein feature information, providing high-resolution protein mass spectra.

Rich Databases and Algorithms

We utilize various protein sequence databases, such as UniProt, Swiss-Prot, etc., and advanced search algorithms such as SEQUEST, Mascot, etc., to achieve high-precision protein identification and mutation analysis.

High-Throughput Data Analysis Platform

We have developed a robust data analysis platform using the latest bioinformatics tools and algorithms to perform various analyses including protein identification, quantification, functional annotation, enrichment analysis, and so on.

Strict Quality Control

We adhere to rigorous quality control processes from sample handling to data analysis, ensuring the services we provide are highly accurate and reproducible.

Personalized Customized Services

We offer customized experimental designs and data analysis solutions to meet specific research objectives of different clients.

Applications

Discovery of New Proteins

Used for designing novel proteins to enhance specific functions or alter their structures to meet specific application requirements.

Study of Protein Structure and Function

Comparing protein sequences and structural differences between different species or individuals can lead to the development of computational methods to predict protein structure and function, furthering understanding of the formation and evolution mechanisms of biodiversity.

Molecular Evolution

Tracing the evolutionary history of organisms can elucidate the evolutionary relationships and functional evolution of different species.

Development of Drugs

Investigating the interactions between proteins and drugs facilitates the development of drugs targeting specific targets.

Diagnosis and Treatment of Disease

Used to identify protein mutations associated with certain diseases, .

The amino acid sequence of proteins forms the basis for determining protein structure and function. Protein sequencing, which determines the types and arrangement of amino acids composing a protein, is a crucial aspect of proteomic studies. Common methods include mass spectrometry (such as tandem mass spectrometry) and the Edman degradation method. Unknown proteins typically refer to those proteins that have not been thoroughly studied and characterized. These proteins may be encoded by newly discovered genes or may be gene products from known organisms whose structure, function, and roles have not been fully understood. Sequencing unknown proteins contributes to revealing the complete picture of an organism's genome and proteome, which is crucial for studying protein structure and function. It also promotes advancements in biotechnology, drug development, and disease diagnosis.MtoZ Biolabs provides integrate Unknown Proteins Sequencing Service.

The MtoZ Biolabs utilizes the latest Thermo Obitrap Exploris 240 mass spectrometer for sequencing unknown protein samples. It employs six commonly used proteases (Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C, and Lys-N) to cleave the target proteins and conduct mass spectrometric analysis. While obtaining fragmented peptide segments, the protein sequence is determined 100% by piecing together these peptide segments.

Analysis Workflow

Sample Preparation

Extraction of proteins and removal of impurities through purification processes.

Protein Fractionation/Isolation

Proteins are fractionated or isolated, typically using one-dimensional or two-dimensional gel electrophoresis (1D/2D-PAGE) and liquid chromatography (such as HPLC).

Protease Digestion

Proteins are digested into smaller peptide segments through multiple enzyme digestions (such as Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C, and Lys-N) to facilitate subsequent mass spectrometric analysis.

Peptide Separation

Peptide segments obtained from protease digestion are further separated using liquid chromatography (such as RP-HPLC).

Mass Spectrometric Analysis

Peptide segments are analyzed using a mass spectrometer to obtain information about their mass, charge states, etc.

Peptide Sequencing

Peptide segments are further sequenced using tandem mass spectrometry (MS/MS).

Data Processing and Analysis

Specialized mass spectrometry software such as Mascot, Sequest, etc., are used to process and analyze mass spectrometry data in combination with known protein databases.

Applications

Discovery of New Proteins

Sequencing and functional annotation of unknown proteins aid in discovering new biological catalysts, signal transduction proteins, transcription factors, and other molecular functions.

Protein Function Studies

Protein sequencing provides data support for functional annotation and protein interaction network studies.

Discovery of Disease Biomarkers

Comparative analysis of proteomes between pathological and normal tissues helps in identifying potential disease biomarkers, providing clues for disease diagnosis and treatment.

Drug Development

Identification of target proteins for drug action and discovery of new drug targets accelerate the development and optimization of new drugs.

Sample Submission Requirements

We accept the following types of samples:

Purified Proteins

Please provide protein samples with purity greater than 85%. The recommended sample volume is 50-200 g, and the protein concentration should be between 0.1-1 mg/mL.

Protein Extracts

Including but not limited to protein extracts from cells, tissues, .

Mass spectrometry-based sequence analysis of proteins, antibodies, peptides, and other samples involves the analysis of the sample's amino acid sequence. Mass spectrometry sequencing refers to the determination of the primary structure of amino acid sequences based on mass spectrometry. Before moving on to mass spectrometry sequence analysis, we provide a brief introduction to a few definitions. Amino acids are organic compounds containing amino (-NH2) and carboxyl (-COOH) functional groups and a side chain (R group) specific to each amino acid. Peptides are short chains composed of two to fifty amino acids linked by peptide bonds. Polypeptides are longer, continuous, unbranched peptide chains, containing up to about fifty amino acids. Peptides/polypeptide chains containing more than fifty amino acids are referred to as proteins. Proteins consist of one or more polypeptides arranged in a biologically functional group manner, typically able to bind to ligands (such as coenzymes and cofactors), another protein, other large molecules (such as DNA or RNA), or more complex macromolecular assemblies. Antibodies (Ab), also known as immunoglobulins (Ig), are large Y-shaped proteins primarily produced by plasma cells, used by the immune system to neutralize pathogens, such as pathogenic bacteria and viruses.MtoZ Biolabs provides integrate Protein Sequencing Service by Mass Spectrometry.

Compared to the Edman degradation method, the advantages of mass spectrometry sequencing are that it is more sensitive, can cleave peptides faster, and can identify terminally blocked or modified protein samples. MtoZ Biolabs utilizes existing high-resolution mass spectrometry technology platforms to provide mass spectrometry-based sequence analysis services, achieving 100% coverage of the targeted protein sequence.

The process of protein sample sequence analysis generally only uses Trypsin for proteolytic digestion of the protein, with peptide coverage of approximately 60%. To obtain the full sequence information of the target protein, MtoZ Biolabs selects six common proteases (Trypsin, Chymotrypsin, Asp-N, Glu-C, Lys-C, and Lys-N) used in the protein sequence analysis process to perform enzymatic digestion and mass spectrometry on the target protein. In the process of obtaining fragmented peptide fragments, the complete protein sequence is determined by splicing the peptide fragments. MtoZ Biolabs uses the Thermo company's newly launched Orbitrap Fusion Lumos mass spectrometer for protein sample sequence analysis. The Orbitrap Fusion Lumos mass spectrometer is currently the highest resolution and sensitivity mass spectrometer, ensuring the sensitivity of identification of low-abundance peptide fragmentation fragments; it also adopts a combination of HCD and ETD modes during the peptide fragmentation process to ensure the integrity of the peptide fragmentation fragments. This can achieve N-terminal, C-terminal sequence analysis, and full-length protein sequence analysis. For samples with an unknown theoretical sequence, de novo sequencing can be performed for sequence analysis.

Mass spectrometry N-terminal sequence analysis is a method that utilizes mass spectrometry technology to efficiently and accurately determine the amino acid sequence of the N-terminus of proteins/peptides. Due to its high sensitivity, high resolution, and high throughput, it can be used to analyze complex protein/peptide mixtures, as well as identify modifications, mutations, and other sequence variations. Compared to traditional Edman degradation, mass spectrometry offers higher throughput and sensitivity, enabling the analysis of more complex samples and larger proteins.MtoZ Biolabs provides integrate MS-Based Protein N-Terminal Sequence Analysis Service.

Based on high-resolution mass spectrometry for N-terminal sequencing, the sample is processed with SDS-PAGE to separate the target protein followed by enzyme digestion. The digested products are analyzed using a Q-Exactive series mass spectrometer (Thermo Fisher Scientific), with peptide fragments in the mass spectrum breaking down into b and y ions' secondary mass spectra. By comparing these spectra with the theoretical amino acid sequence of the protein, the N-terminal position of the protein is determined. MtoZ Biolabs combines high-performance liquid chromatography and high-resolution mass spectrometry, based on the principle of N-terminal sequencing, to establish a mass spectrometry-based N-terminal sequence analysis platform, providing you with high-quality mass spectrometry N-terminal sequence analysis services. We use two different proteolytic cleavage methods to produce two different lengths of N-terminal peptide segments, which are mutually verified to finally determine the protein N-terminal sequence. Welcome your consult for more detailed information!

Experimental Instruments

High-performance Liquid Chromatography Tandem Mass Spectrometer: High-performance Liquid Chromatograph-Easy-nLC 1200Electrospray-hybrid Ion Trap Orbitrap Mass Spectrometer - Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer

Service Advantages

High-resolution Mass Spectrometry-Based N-terminal Sequencing for Analysis of N-terminal Capping and Post-translational Modifications (PTMs), Determining the Protein N-terminal Starting PointSupport for N-terminal Dimethylation Labeling in Complex Proteins by our Company, Involving Analysis of Labeling Positions to Determine the N-terminal Starting Points in Complex Protein Samples

Case Study

N-terminal Amino Acid Sequence Analysis of a Protein Drug Sample:

After trypsin digestion, the sample is analyzed by LC-MS/MS, and the mass spectrometry data is matched with a theoretical database, obtaining an MS2 spectrum consistent with the theoretical N-terminal sequence (HYAHVDCPGHADYVK), as shown below:

From the analysis above, it can be concluded that the N-terminal starting amino acid of this protein drug is histidine, with the N-terminal sequence as HYAHVDCPGHADYVK.

Services at MtoZ Biolabs

Experimental ProceduresMass Spectrometric ParametersDetailed Information on Mass Spectrometry N-terminal Sequence AnalysisMass Spectrometric ImagesRaw Data

Accurate mass determination can be applied to all areas involved in chemistry and biochemistry. It is used to identify unknown compounds in complex mixtures, thereby simplifying the identification of all components. The accuracy of the determination refers to the degree of proximity between the measured value and its true value. In recent years, the mass determination of biological samples has received increasing attention, and at the same time, accurately determining the mass of these compounds has faced new challenges. Various instruments, especially advancements in MS, have made the application of accurate mass determination through MS more widespread.MtoZ Biolabs provides integrate Accurate Mass Determination Service.

MtoZ Biolabs offers precise mass determination services for proteins, oligonucleotides, and oligosaccharides. MtoZ Biolabs mass spectrometry laboratory ensures more reliable and accurate determination results by regularly calibrating instruments, maintaining equipment, and establishing standard curves.

Sample Submission Requirements

We recommend using high quality solid samples or solutions. If you want to know the specific requirements for special samples, welcome to contact us.

MtoZ Biolabs uses the most advanced mass spectrometer-the Thermo Scientific Orbitrap Fusion Lumos-coupled with nanoLC-MS/MS nano-scale chromatography, to perform accurate mass determinations with high resolution and precision.

Servie at MtoZ Biolabs

Sample Processing ProcedureExperimental Methods and StepsRaw DataData Analysis Report

Protein tandem mass spectrometry (MS/MS) is an efficient, precise, and sensitive method for qualitative and quantitative identification of proteins based on mass spectrometry technology. It is widely used in proteomics research, biopharmaceutical quality control, disease biomarker discovery, and other fields. By analyzing the peptide fragments (products of protein digestion) using mass spectrometry, this technique identifies target proteins, including unknown ones. Originating in the early 1990s, it was initially dominated by liquid chromatography coupled with mass spectrometry (LC-MS/MS). With the development of techniques such as matrix-assisted laser desorption/ionization (MALDI), protein mass spectrometry identification entered a new era. Today, MS/MS has become an important tool in proteomics research, with significant implications for deciphering protein structures and functions in complex biological samples. It aids in uncovering protein interactions, signal transduction mechanisms, and drives advancements in biological science and disease treatment.MtoZ Biolabs provides integrate Protein Identification Service by Tandem Mass Spectrometry.

Analysis Workflow

Sample Preparation

Purify, concentrate, and desalt the provided protein samples from customers using appropriate methods such as high-performance liquid chromatography (HPLC) or gel electrophoresis to remove impurities and enhance protein purity.

Protein Digestion

Perform multiple enzymatic digestions on purified proteins to cleave them into peptide fragments, facilitating subsequent mass spectrometry analysis.

Peptide Separation

Separate peptide fragments using nano-high-performance liquid chromatography (nano-HPLC) to enhance resolution and detection sensitivity effectively.

MS/MS Analysis

Employ high-resolution, high-sensitivity mass spectrometers (Thermo Fisher's Orbitrap Fusion Lumos, Orbitrap Exploris 240) to analyze the quality and fragment ions of separated peptide fragments through MS/MS.

Data Analysis

Utilize specialized data processing software such as Mascot and protein databases for peptide quality matching and fragment ion searching, ultimately achieving protein identification, including detailed information such as protein name, amino acid sequence, and matching score.

Applications

High-Throughput Protein Identification

MS/MS plays a critical role in proteomics research, identifying and quantifying proteins in biological samples, revealing differences in protein expression, interactions, and functions.

Post-translational Modification Studies

Used for identifying post-translational modifications of proteins (such as phosphorylation, acetylation, glycosylation), revealing the impact of modifications on protein function and signaling.

Protein Interaction Studies

By identifying components of protein complexes, it reveals the interaction relationships between proteins, providing a basis for the study of biological signaling pathways.

Protein Structure Analysis

MS/MS technology can analyze the primary structure (amino acid sequence) and some secondary and tertiary structure information of proteins, providing reference data for protein structure research.

Disease Diagnosis and Biomarker Research

MS/MS technology can be used for the identification and quantification of disease-related proteins, providing a basis for disease diagnosis and prognosis assessment.

Sample Submission Requirements

Deliverables

In the technical report, MtoZ Biolabs will provide you with a detailed report including:

Interacting protein mass spectrometry identification is a method utilizing mass spectrometry technology to study protein-protein interactions. It separates target protein complexes through methods such as immunoprecipitation (IP), affinity purification (AP), etc. Then, it employs protein enzymatic digestion to generate peptide fragments, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of these peptides. Finally, based on the mass spectrometry data, combined with bioinformatics tools, it identifies and quantifies interacting proteins. Interacting protein mass spectrometry identification can identify or verify interacting proteins, discover new proteins interacting with target proteins, identify and quantify interacting proteins, and elucidate protein signaling pathways in organisms, aiding in drug target discovery and biomarker research, and providing a basis for drug development.MtoZ Biolabs provides integrate MS-Based Protein Interaction Identification Service.

MtoZ Biolabs provides one-stop interacting protein mass spectrometry identification services based on Thermo Fisher's Orbitrap Exploris 240 mass spectrometry platform combined with Nano-LC chromatography. This service analyzes the mass spectrometry identification of proteins/protein mixtures in IP, Co-IP samples, GST fusion protein Pull-down, and other purified samples. The protein sample pre-processing in the identification experiment strictly follows the article published in Nature Methods: "Universal sample preparation method for proteome analysis, " ensuring higher peptide coverage in the identification experiment.

Analysis Workflow

Protein Extraction

Optimal methods are employed to extract proteins from cell or tissue samples, and the purity and concentration of proteins are determined to ensure the quality of the extract.

Interacting Protein Acquisition

Target proteins and their interacting proteins are enriched using IP, GST-Pull-down, or immunoblotting methods according to experimental requirements.

Elution and Enrichment

Target protein complexes are eluted from the binding medium, enriched, and protein samples are collected for the subsequent mass spectrometry analysis.

Protein Digestion

The eluted interacting protein complex samples are enzymatically digested with various proteases (such as trypsin) to generate peptide fragments.

Peptide Separation and Purification

Peptide fragments obtained after digestion are separated and purified using high-performance liquid chromatography (HPLC) or reversed-phase high-performance liquid chromatography (RP-HPLC) techniques.

Mass Spectrometry Analysis

Peptide fragments are subjected to mass spectrometry analysis via LC-MS/MS to obtain mass spectrometry data.

Data Processing and Analysis

Mass spectrometry data is subjected to database searches and bioinformatics analysis for qualitative and quantitative identification of interacting proteins.

Service Advantages

High Sensitivity and Accuracy

Our mass spectrometry technology accurately detects interacting proteins even in complex biological samples, including low-abundance interacting proteins, with high sensitivity.

High Throughput

Mass spectrometry technology can identify and quantify a large number of proteins in a short time, saving researchers time and experimental costs.

Diverse Protein Interaction Types

Our services cover various protein interaction types, including homologous interaction, heterologous interaction, transient interaction, permanent interaction, and among others.

Relative Unbiasedness

Proteins serve as the carriers of life substances and executors of biological functions, playing a crucial role in maintaining normal physiological activities of organisms. Protein characterization analysis involves the characterization of protein purity, molecular weight, structure, post-translational modifications, and interactions, among others. As one of the core areas in biological science research, protein characterization analysis reveals how proteins function within biological systems and how they interact to maintain normal physiological functions.

Common methods for protein characterization analysis include nuclear magnetic resonance (NMR), mass spectrometry (MS), and Western blotting (WB). NMR is commonly used to study the three-dimensional structure and dynamic properties of proteins. MS analyze the molecular weight, post-translational modifications, and interactions of proteins. While WB utilizes specific antibodies to detect distances and interactions between proteins. MtoZ Biolabs provides integrate Protein Characterization Service.

Services at MtoZ Biolabs

Protein Purity Analysis

Various purity analysis methods are available, including SDS-PAGE, capillary electrophoresis, zinc reverse staining, glutaraldehyde-modified silver staining, and MALDI-TOF-MS. These methods are utilized to detect contaminants, protein variants, isoforms, S-S bond mismatches, truncated proteins, degraded proteins, protein modifications, protein aggregates, and protein precursors.

Protein Molecular Weight Analysis

Protein complexes and protein-small molecule complex molecular weights are determined using techniques such as gel permeation chromatography, SDS-PAGE, and mass spectrometry.

Protein Structure Identification

Characterization of top-down protein primary structure (including amino acid composition analysis, peptide sequence analysis, and disulfide bond localization) is performed based on Thermo Fisher's Orbitrap Fusion Lumos mass spectrometry platform combined with nanoLC-MS/MS chromatography, as well as protein spatial configuration determination based on circular dichroism spectroscopy.

Protein Post-Translational Modification Analysis

Multiple mass spectrometry platforms (Thermo Fisher's Q Exactive HF, Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometry platforms combined with Nano-LC) are employed for the identification of protein phosphorylation, glycosylation, ubiquitination, acetylation, methylation, disulfide bond formation, nitrosylation, and among other post-translational modifications.

Protein Interaction Analysis and Identification of Interacting Proteins

Techniques such as WB, immunoprecipitation (IP), and GST-pull-down are utilized to study protein-protein interactions. LC-MS/MS is employed for mass spectrometric identification analysis of proteins/protein complexes in IP, Co-IP samples, and GST fusion protein pull-down purified samples.

Deliverables

In the technical report, MtoZ Biolabs provides detailed technical reports, including:

Experimental ProceduresRelevant Mass Spectrometry ParametersDetailed information on protein characterization analysisMass Spectrometry ImagesRaw Data

Ideally, any technology for detecting Host Cell Protein (HCP) should: (i) detect protein concentrations across a wide dynamic range, from extremely low concentrations of individual impurity proteins to much higher concentrations of impurity proteins; (ii) track changes in HCP populations and concentrations throughout the entire bioprocess; (iii) simultaneously monitor and measure multiple protein analytes; and (iv) monitor and measure low concentration HCP even in the presence of target recombinant protein at high concentrations. Additionally, if a risk-based assessment of HCP is necessary, these methods should be capable of identifying HCP at any stage of the bioprocess. Employing anti-HCP antibodies for qualitative and quantitative detection and removal of HCPs is highly effective. Thus, the development of anti-HCP antibodies is of significant importance.

1. Identification of HCP Antigens

A variety of cell lines, including mammalian, insect, plant, and prokaryotic cells such as Escherichia coli, are employed for producing recombinant proteins for therapeutic purposes. Chinese hamster ovary cells (CHO) are widely used in the biotechnology industry. For instance, pIs range from 3-11, with molecular weights from 5kDa to 250kDa for large proteins. During the purification process, the content of the product protein gradually increases, while the content of host impurity proteins significantly decreases, dropping from >10-90% to.

About Us

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

Email: marketing@mtoz-biolabs.com

Peptide mass fingerprinting (PMF), also termed protein fingerprinting, is a high-throughput analytical technique developed in 1933 for protein identification. It involves the cleavage of an unknown target protein into smaller peptides by endopeptidases, followed by the precise measurement of these peptides' absolute mass using a mass spectrometer to generate a peptide peak list. This list is then compared against a theoretical peptide mass database using bioinformatics tools. This database, which contains translated proteins from all known genomes, also predicts peptide masses post-hydrolysis, facilitating the identification process through statistical comparison of experimental peptide masses with theoretical values. MtoZ Biolabs provides integrate Peptide Mass Fingerprinting Analysis Service.

Analysis Workflow

Proteins isolated from biological tissues or cells are separated via 2D or SDS-PAGE and analyzed using imaging techniques. Proteins showing significant differences are selected for further analysis, which involves gel extraction and enzymatic digestion to yield peptides that characterize the protein. The peptide mixture is then analyzed through mass spectrometry to obtain a detailed fingerprint spectrum, which aids in molecular weight determination and facilitates database matching.

Applications

PMF has gained widespread application across various domains, including drug authenticity verification, food quality control, and disease diagnosis, due to its high sensitivity, robustness, and minimal sample requirements. Recognized by the World Health Organization, it serves as a critical technique for material identification and quality control.

Service Advantages

Protein Identification Relies on Peptide Mass rather than SequencingClassical Method Employing First-Stage Mass Spectrometry for Rapid Protein Identification

Limitations

The Sequence of the Target Protein Must be Present in the DatabasePrimarily Applicable to Single Protein Analysis, the Presence of Mixed Proteins Complicates the AnalysisSimilar Peptide Masses Increase the Challenge of Accurate Matching

MALDI-TOF PMF

Rapid and precise protein identification is essential in proteomics. PMF employs matrix-assisted laser desorption/ionization (MALDI) coupled with time-of-flight (TOF) mass spectrometry, making it a prevalent choice.

MALDI uses a laser to irradiate the eutectic film formed by the sample and substrate. The matrix absorbs energy from the laser and transfers it to the biomolecules. During the ionization process, protons are transferred to or from biomolecules, causing ionization. Thus, MALDI is a soft ionization technique, suitable for the determination of mixtures and biological macromolecules. The principle of TOF is that ions are accelerated through a flight tube under the influence of an electric field, and ions are detected according to the time it takes to fly to the detector. MALDI-TOF-MS has high sensitivity, precision, and resolution. It plays a crucial role in sample analysis in life sciences and many other fields.

MtoZ Biolabs provides specialized protein identification services utilizing peptide fingerprinting PMF technology, and welcomes your inquiries.

Deliverables

In the technical report, MtoZ Biolabs will provide you with a detailed technical report, including:

Experimental ProceduresRelevant Mass Spectrometry ParametersMass Spectrometry ImagesRaw DataResults of Protein Molecular Weight and Purity Analysis

Proteins exhibit asymmetric secondary structures such as -helices, -sheets, -turns, and others, leading to differential absorption of left and right circularly polarized light. Upon transmission through these proteins, the circularly polarized light becomes elliptically polarized, a phenomenon known as protein circular dichroism (CD). CD Spectroscopy is an essential technique for analyzing the secondary and tertiary structures of optically active molecules (e.g., proteins, DNA) in solution. It is widely utilized in studying the structure-function relationship and interactions of macromolecules. MtoZ Biolabs provides integrate Protein Circular Dichroism Analysis Service.

MtoZ Biolabs offers professional CD services for analyzing and determining the spatial configurations of proteins.

Service Advantages

Shorter, Simpler, and More Efficient Measurement ProceduresRequires Minimal Sample Volume and Can be Performed in Dilute SolutionsNo Limitations Regarding Molecular Weight or SizeHighly Sensitive to Changes in Secondary and Tertiary Structures, Capable of Detecting Subtle Alterations

Figure 1. CD Spectroscopy Illustrates the Representative Secondary Structures of Peptides and Proteins

Sample Submission Requirements

Protein sample concentration should exceed 0.5 mg/mL, with purity exceeding 90%. The minimum quantity of solid protein should not be less than 200 g.

Deliverables

In the technical report, MtoZ Biolabs will provide you with a detailed technical information, including:

Experimental ProceduresRelevant Data

Protein isoform analysis is a method to study the differences in structure and function of proteins. Proteins are key molecules in life activities, and their functions are closely related to their structures. However, the same amino acid sequence may form multiple different three-dimensional structures, which are called isoforms. Protein isoforms may result from various factors, such as post-translational modifications, heterologous expression, environmental factors, etc. Isoformization of monoclonal antibodies (mAbs) is one of the common pathways of protein degradation. Isoforms can not only cause changes in protein function but are also associated with various diseases, such as neurodegenerative diseases, cancer, etc. Therefore, the analysis and identification of protein isoforms are crucial, especially in drug research and development and protein drug production. MtoZ Biolabs provides integrate CE-SDS Protein Isoform Analysis Service.

Capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) technique is effective in detecting and differentiating protein isoforms. Due to the differences in charge, size, and shape among protein isoforms, they migrate at different rates in an electric field, allowing the separation, detection, and identification of these isoforms by CE-SDS. The advantage of this technique lies in its ability to analyze both purified protein samples and complex biological samples, such as cell lysates or tissue extracts. The high resolution of CE-SDS enables it to distinguish isoforms with very similar structure. It provide a powerful tool for the structural and functional study of protein isoforms with high sensitivity, resolution, and speed.

MtoZ Biolabs has developed and validated a protein isoform CE-SDS analysis platform based on high-precision capillary electrophoresis instrumentation and protein electrophoresis principles, offering CE-SDS protein isoform analysis services. This service can be used to analyze protein products such as monoclonal monomers under normal and stressful conditions, aiding in the early identification of degradable drug candidates during drug development.

Service at MtoZ Biolabs

Experimental ProceduresRelevant Experimental ParametersDetailed Information on Protein IsoformsElectrophoresis ImagesRaw Data

The term "proteomics" is the combination of "protein" and "genomics", referring to the full set of proteins expressed by a genome, which consists of all the proteins expressed by a cell or an organism. In 1995 (though there are mentions of 1994 and 1996), Marc Wilkins first proposed the concept of proteome, and in 1997, Peter James (who worked at the Swiss Federal Institute of Technology, known as the European MIT) further introduced the concept of proteomics. The concepts of genomics and proteomics have further given rise to various other omics, and their birth and development have also made systematic biology possible. MtoZ Biolabs provides integrate Proteomics Analysis Service.

Research on proteome not only provides a material basis for understanding the laws of life activities but also offers theoretical basis and solutions for elucidating numerous disease mechanisms. By comparing the proteome of normal and pathological individuals, we can identify certain disease-specific protein molecules, which may serve as molecular targets for new drug design, or may also provide molecular markers for early disease diagnosis. Indeed, some of the best-selling drugs worldwide are either proteins or target-specific protein. Thus, proteomics research is not only essential for exploring the mysteries of life but also benificial to human health. The study of proteomics is a characteristic of life sciences entering the post-genomic era.

Figure 1. Applications of Proteomics

Proteomics refers to the systematic identification and quantification of the proteome of a biological system (cell, tissue, organ, body fluid, or organism) at a specific time point. Mass spectrometry (MS) is the most commonly used technique for proteomics. MtoZ Biolabs, based on high-throughput mass spectrometry technology, has established many proteomics analysis platforms, possessing first-class MS-based proteomics techniques and analysis capabilities, to provide a complete set of proteomics solutions.

Protein Identification

Protein identification is a fundamental step in proteomics, aimed at identifying the proteins in a sample, and is critical for determining the nature of protein products. The molecular weight, isoelectric point, structure, and amino acid sequence of a protein are key attributes in determining the type of protein. MtoZ Biolabs possesses high-resolution mass spectrometry, capillary electrophoresis, and circular dichroism methods for protein identification, enabling precise identification of protein by detecting these attributes and meeting various research needs. Whether your sample is protein mixture, SDS-PAGE protein bands, 2D protein gel spots, or pull-down and Co-IP samples, we can provide efficient and accurate identification.

Quantitative Proteomics Analysis

Protein quantification is the process of determining the relative or absolute quantity of proteins in samples. In proteomics research, researchers often need to compare protein expression levels under different conditions, such as normal versus disease states, or research changes of protein expression levels over different time points. Quantification of protein can be achieved through various methods, such as isotopic labeling (e.g., TMT, iTRAQ, SILAC, 15N labeling) and label-free quantification, providing information on protein quantities in different biological samples. MtoZ Biolabs quantitative proteomics platform offers various protein quantification techniques, including Label-free, iTRAQ, TMT, SILAC, 2D-DIGE, DIA, SWATH, and relative, semi-quantitative, .

AQUA absolute quantitative analysis is a targeted quantitative proteomics technique widely used in various quantitative proteomics studies. The absolute quantification strategy can be used for the absolute quantification of proteins and their modified states. By using stable isotope-labeled synthetic peptides as internal standards, it can simulate the natural peptides formed by protein hydrolysis, and can also be prepared using covalent modifications (such as phosphorylation, methylation, acetylation, etc.). By using selected reaction monitoring (SRM) analysis in MS/MS, AQUA internal standard peptides are used for precise quantitative determination of the absolute levels of proteins and post-translationally modified proteins after protein hydrolysis. MtoZ Biolabs provides integrate Absolute Quantitative Analysis (AQUA) Service.

The advancement of MS technology has promoted the development of large-scale quantitative analysis methods for intracellular protein expression levels.

Analysis Workflow

Figure 1. Analysis Workflow of AQUA

Protein absolute quantification strategy is an effective detection method for protein quantification and post-translational modifications. Absolute quantification is achieved by using synthetic peptides labeled with stable isotopes: based on discovering the relationship between mass spectrometry signal and protein concentration, the average mass spectrometry signal response of the three strongest trypsin peptides per mole of protein is constant, with a coefficient of variation less than 10%. As long as a given internal standard (a synthetic peptide containing isotopic labels) is provided, the relationship between mass spectrometry signal and protein concentration can be used to determine a universal signal response factor (counts/mol). The universal signal response factors for all quantified proteins are the same.

The absolute quantification method relies on the use of synthetic internal standard peptides, which are introduced into the cell lysate at a known concentration during the digestion process. During the detection process in a tandem mass spectrometer, the hydrolyzed protein samples are analyzed by the SRM method, allowing for direct detection and quantification of natural peptides and isotopically labeled AQUA internal standard peptides. This method's simplicity and sensitivity, combined with the widespread use of tandem mass spectrometers, make the AQUA absolute quantification strategy an effective method for directly measuring protein levels and post-translational modifications from cell lysates.

The process where proteins separated by electrophoresis are transferred or blotted from the gel matrix onto a membrane (usually nitrocellulose or polyvinylidene difluoride (PVDF)), followed by subsequent antibody-based detection on the membrane surface, is known as western blot (WB) or immunoblotting. Mtoz Biolabs provides WB analysis services. Through highly selective and sensitive antibody-antigen interactions, WB can be used to detect specific target proteins from complex protein mixtures, such as tissue homogenates or cell extracts. The obtained data can be used for qualitative and semi-quantitative analysis of target proteins. MtoZ Biolabs provides integrate Protein Immunoblotting and Electrotransfer Services.

Analysis Workflow

Figure 1. General Flowchart of Electric Transfer

WB is a widely used protein analysis technique for detecting specific proteins in tissue homogenates or extract samples. It uses gel electrophoresis to separate native proteins and denatured proteins by their 3D structures and peptide chain lengths, respectively. After separation, the proteins are transferred onto a membrane, where specific antibodies recognize the target proteins specifically. WB is widely applied in molecular biology, biochemistry, immunogenetics, and other molecular biology fields.

In WB analysis, proteins are first separated from gel electrophoresis by various methods such as SDS-PAGE and IEF, by isoelectric point (pI), molecular weight, charge, or a combination of these factors. SDS-PAGE is commonly used for separation because all proteins are dissolved within the gel and migrate in the same direction, and the denaturing effect of SDS makes antigenic epitopes more easily recognizable.

After gel electrophoresis analysis, proteins are transferred from the gel to a membrane made of nitrocellulose or PVDF for antibody detection. PVDF membranes have higher protein binding capacity than nitrocellulose membranes, but nitrocellulose membranes are better at binding smaller proteins. The main method of transferring proteins is electroblotting, which uses electricity to pull proteins from the gel into the PVDF or nitrocellulose membrane. Electrophoretic transfer methods include wet transfer, semi-dry transfer, and semi-wet transfer. If protein separation is done by IEF, then using pressure transfer diffusion to transfer proteins is more effective.

The transfer process varies from one hour (semi-dry transfer) to overnight transfer (wet transfer). After the transfer is complete, the free binding sites on the membrane are blocked by the protein mixture, thus subsequent antibody detection is not interfered. The proteins transferred to the membrane can then undergo antibody detection. Membranes containing proteins are first incubated with primary antibodies, followed by detection with secondary antibodies that recognize the primary antibodies. Secondary antibodies generally carry a reporter group and have high sensitivity.

The most sensitive detection method is using enhanced chemiluminescence (ECL): recognizing primary antibodies with an antibody-horseradish conjugate. Using special variants of ECL, protein bands as low as 1 pg can be detected.

WB can also be used to track the phosphorylation of proteins, with antibodies that bind to all subtypes of phosphorylated proteins presenting a bead-like appearance on 2D gels. It can also be used to identify known and unknown proteins in complexes produced by IP, as long as the target proteins are in the gel, they can be identified by coomassie brilliant blue staining, .

Multiple reaction monitoring (MRM) is a targeted quantitative proteomics research method for target protein. MRM, based on information about the target molecule, selectively chooses data for mass spectrometry data collection, collecting signals that meet the criteria of target ions and eliminating that of ions that do not meet the criteria. MRM mass spectrometry analysis goes through three stages: 1) filtering out parent ions consistent with the specificity of the target molecule through MS; 2) colliding and fragmenting these parent ions to eliminate interference from other ions; 3) collecting mass spectrometric signals from selected specific MS/MS2 ions only. MRM mass spectrometry technology is a high-precision protein quantification technique and is an excellent method for one-time precise quantification of multiple target proteins in complex samples. If isotope-labeled target peptides are used as internal references, absolute quantification of proteins can be achieved. MtoZ Biolabs provides integrate MRM/PRM Quantitative Proteomics Service.

MtoZ Biolabs utilizes AB SCIEX TripleTOF 5600, AB SCIEX Triple Quad 5500, Q Exactive, and Fusion mass spectrometry platforms combined with Nano-LC, launching MRM/PRM targeted quantitative proteomics analysis service. You just need to provide us with the information about the target proteins you need to study, and we provide a one-stop MRM quantitative proteomics analysis service. It includes establishing and optimizing MRM methods, selecting specific peptides, labelling peptides by isotope, analyzing by mass spectrometry, analyzing raw data, and analyzing by bioinformatics method.

Experimental Instruments

AB SCIEX TripleTOF 5600, AB SCIEX Triple Quad 5500, Q Exactive, Fusion

Analysis Workflow

Service Advantages

High sensitivity, selecting of ions consistent with the target ions by two-stage mass spectrometry and excluding interfering ions to greatly enhance the signal-to-noise ratio and improve the accuracy of detected target ions.High throughput, the number of protein identified can reach up to 200 at once.Capablility of absolute quantitative analysis without antibodies.High-precision identification of low-abundance proteins and quantitative range across four orders of magnitude.

Applications

Application of MRM Technology

(1) Result validation of label-free and other non-targeted proteomics.

(2) Simultaneous absolute quantification study of multiple proteins/peptides.

(3) Study the change of protein families with high homology but lacking specific recognition antibodies.

(4) Quantitative study of post-translational modifications of proteins.

(5) Absolute quantification study of biological disease targets.

Application of SRM/MRM

(1) Verification of iTRAQ differential proteins.

(2) Verification of label-free differential protein post-products.

(3) Absolute quantification of peptides and proteins.

(4) Quantification of disease markers and establishment of diagnostic models.

(5) Quantification of phosphorylated proteins and methylated proteins.

(6) Quantification of other post-translational modifications protein.

(7) Quantitative analysis of pathway .

Sample Submission Requirements

Deliverables

MRM Spectrum Analysis and Protein Data Quality AssessmentPCA of Multi-component SamplesIdentified Protein Functional Annotation: GO Functional Annotation, KEGG Functional Annotation and COG Functional AnnotationDifferential Protein Expression Statistical Analysis: Venn Diagrams and Volcano PlotsDifferential Protein Expression Clustering Analysis:.

4D-DIA quantitative proteomics is an emerging high-throughput mass spectrometry technology that combines data-independent acquisition (DIA) strategies with four-dimensional (4D) separation techniques. It involves digesting protein into peptide by protease and separating peptide by liquid chromatography (LC), followed by mass spectrometry (MS) for detection and identification of peptide. It enables high-throughput and high-sensitivity quantitative analysis of proteins in biological samples. 4D-DIA is widely used in fields such as biomedicine research, disease diagnosis, and drug development. Its in-depth analysis of proteome helps reveal biological process, discover potential biomarker, and study drug targets to promote scientific research and clinical applications. MtoZ Biolabs provides integrate 4D-DIA Quantitative Proteomics Service.

Analysis Workflow

Sample Preparation

Proteins extraction from biological samples, followed by purification and concentration.

Protein Digestion

Protein digestion with enzymes (such as trypsin) to produce peptides.

Peptide Separation

Peptides separation by high-performance liquid chromatography (HPLC).

Mass Spectrometry Analysis

4D-DIA analysis of separated peptides by mass spectrometer, including their retention time (RT), mass-to-charge ratio (m/z), ion intensity, and ion mobility, followed by mass spectrometry data collection and mass spectrum match.

Data Processing and Quantitative Analysis

Identification of peptide and quantification of protein by searching for mass spectrometry databases.

Service Advantages

High Throughput

4D-DIA technology can analyze a large number of samples simultaneously, significantly reducing experimental cycle and improving experimental efficiency.

High Accuracy

4D-DIA technology uses high-sensitivity, high-resolution mass spectrometers for protein quantification, offering high accuracy and reliability and avoiding error and omission.

Comprehensive Coverage

4D-DIA technology can analyze almost all proteins in samples, not just specific ones.

Data Reproducibility

The data generated by 4D-DIA technology have high repeatability and reproducibility, ensuring the stability and credibility of experimental results.

Applications

Functional Proteomics Research

Exploration of the structure, function, and interactions of proteins, providing a critical theoretical foundation for biological science.

Protein Modification Research

Modifications of protein can affect its function and regulatory roles. 4D-DIA quantitative proteomics is used to study various types of protein modifications, such as protein phosphorylation, protein acetylation, protein methylation, etc.

Disease Biomarker Identification

Identification of potential disease biomarkers by comparing pathological samples with normal samples, enhancing the accuracy of early diagnosis and efficacy of disease treatment.

New Drug Development

In-depth research on the mechanisms of drug action to develope more targeted medications and offer personalized treatments.

Sample Submission Requirements

We accept a variety of samples, including cell samples, tissue samples, body fluid samples (such as serum, plasma, urine, saliva, etc.), and microorganism samples (such as bacteria, viruses, fungi, etc.).

Service atMtoZ labs

Experimental ProceduresRelevant Mass Spectrometry ParametersDetailed Information on 4D-DIA Quantitative ProteomicsMass Spectrometry ImagesRaw Data

With the increasing focus on personalized healthcare, there is a growing demand for biomarkers for early diagnosis, prognosis, patient stratification, or monitoring treatment response. Compared with genetic biomarkers, protein biomarkers provide higher degrees of differential information. Olink proteomics is based on a unique proximity extension assay (PEA) technique that enables simultaneous quantification of multiple protein biomarkers. Olink PEA uses 2 matching antibodies for each target antigen. Each antibody pair is labeled with a unique DNA oligonucleotide barcode. When antibodies bind to the same target protein in solution, the DNA oligochain will anneal to sufficient stability to achieve DNA elongation. Then the DNA barcode is amplified and the amplicons obtained through qPCR or next-generation sequencing (NGS) is measured for absolute or relative quantification. PEA can detect protein with high sensitivity. Mtoz Biolabs provides integrate Olink Analysis Service.

Olink proteomics provides high-throughput solutions for protein biomarker discovery, enabling the detection of numerous proteins in biological samples with concentration low to microscale, and its analytical methods are rigorously validated, which can provide superior specificity at multiple levels. Due to its high sensitivity and high throughput, Olink proteomics has been widely used in the fields of biomarker discovery, disease mechanism research, drug development and personalized healthcare. In clinical biomarker research, it can be used in all stages of drug development, including early detection and preclinical and clinical development.

Analysis Workflow

Minimum Clinical Sample Concentration (Plasma/Serum as Low as 1 L)High Sensitivity to Protein Detection, Similar to ELISA or BetterImproved Accuracy of Multiple Detection and Intermediate Multiple Detection (with the Average Internal Coefficient of Variation (%CV) Lower than 10% and the Average Intermediate Coefficient of Variation (%CV) Lower than 20%)More than 10 Logarithms of the Dynamic Range Which Is Ideal for Study of Plasma Protein GroupHigh Specificity of Protein Detection Aouble Recognition of Two Antibodies and Unique DNA Barcode of each Protein without Cross-reactivityvailablethrough D

Applications

Drug Target Identification

(1) Identify protein quantitative trait loci (pQTLs) to link genetic variations, proteins, and diseases.

(2) Delve into tumor biology, immune regulation mechanism, phenotypic change, and the transition from cold tumor to hot tumor.

Biomarker Analysis and Screening

Analyze longitudinal biomarker data between early and late time points to identify key biomarkers related to the transition from drug sensitivity to drug resistance.

Protein Response Validation in Clinical Trials

(1) Predict and monitor therapeutic response, sensitivity, and immune resistance mechanism.

(2) Guide strategies to optimize response rates and identify new target pathways.

Exploratory Endpoints in Clinical Trials

(1) Explore the correlation between circulating protein biomarker and clinical outcome.

(2) Predict the onset and mechanisms of treatment-related toxicity and monitor cytokine release syndrome and neurotoxicity.

(3) Predict relapse and treatment progression in adjunct therapy.

Deliverables

In the results report, MtoZ Biolabs will provide you with a detailed results report, which includes:

Experimental ProceduresAbsolute Quantification Data (pg/mL) (Limited to Olink Target 48)Relative Quantification Data: Expressed in Normalized Protein Expression.

Peptidomics refers to the study of all endogenous bioactive peptides in organism, cell, or tissue. Bioactive peptides are biologically active substances involved in various cellular functions within organism, which include cytokines, growth hormones, and disease-specific degradation fragments of certain proteins in body fluid. These peptides play a crucial role in the regulation of organism, encompassing hormone regulation, neurotransmitter modulation, cell growth and proliferation, and immune modulation. The investigation of peptide structures and physiological functions holds significant importance in life sciences. Peptidomics is the discipline that studies the structure and function of the peptidome from multiple perspectives. Mass spectrometry (MS) employed in peptidomics enable both qualitative and quantitative identification of peptide in samples. MtoZ Biolabs provides integrate Peptidomics Analysis Service.

Service at MtoZ Biolabs

Peptide Purity AnalysisPeptidomics AnalysisPeptide Biomarker IdentificationPeptide Mass Spectrometry IdentificationHigh-accuracy MS-based Immunopeptidomics Analysis and Neoantigen DiscoveryPeptide Structure Determination

High performance liquid chromatography (HPLC) is a modern chromatographic separation technique extensively utilized in biological, chemical, and medical research. For peptide purity analysis, HPLC identifies the composition and content of peptides in samples, assessing the purity and impurities. Peptides are dissolved in a mobile phase and separated using a chromatographic column containing a stationary phase. Different components in the peptide sample exhibit distinct retention times based on their interactions. The detector analyzes retention time and signal intensity to qualitatively and quantitatively assess peptide purity. MtoZ Biolabs provides integrate HPLC Peptide Purity Analysis Service.

Analysis Workflow

Sample Preparation

Dissolve peptide samples in an appropriate solvent (e.g., water, acetonitrile, methanol) to form the test solution.

Set HPLC Conditions

Adjust parameters like mobile phase, flow rate, gradient program, and detection wavelength according to sample characteristics and the chromatographic column.

System Equilibration

Flow the mobile phase through the column until reaching a stable equilibrium to ensure reproducible results.

Sample Injection

Introduce the test solution to the chromatography system using an automatic or manual injector.

Separation Process

Peptide samples are carried through the column by the mobile phase, separating components based on their interaction with the stationary phase.

Detection and Data Collection

The detector sequentially registers each peptide component passing through, producing electrical signals. The system captures retention time and signal intensity to create chromatograms.

Data Processing and Analysis

Specialized software processes chromatograms, providing qualitative and quantitative peptide purity analysis based on retention time and signal intensity.

Drug Development and Preparation

Assess peptide drug purity, composition, impurities, and concentration to ensure drug quality and safety while enabling high-purity peptide preparation.

Industrial Production

Monitor purification steps, degradation, and impurity production during peptide manufacturing to guide process optimization and quality control.

Clinical Diagnosis

Measure peptide levels in biological samples (e.g., serum, urine) to support clinical diagnosis, disease monitoring, and treatment evaluation.

Food Safety

Confirm peptide additive and residue purity in food, ensuring safety and quality.

Environmental Monitoring

Detect peptide pollutants in environmental samples (e.g., water, soil) to aid environmental risk assessment and pollution control.

Sample Submission Requirements

In the technical report, MtoZ Biolabs will provide you with a detailed technical information, including:

Experimental ProceduresRelevant Chromatographic ParametersHPLC Peptide Purity Analysis Detailed InformationChromatography ImagesRaw Data

Peptides are abundant in living organisms, actively participating in key biological processes such as hormone regulation, enzyme catalysis, and immune responses. Their spatial structure and functional features greatly influence their biological activity. Accurately determining peptide structures is essential for understanding peptide folding patterns, bioactivity, and structural features. It also informs drug development and bioengineering optimization. Peptide structure determination is valuable in biomedical research and drug development, providing insight into protein structure-function relationships, supporting drug design, and advancing vaccine research. Furthermore, it is critical in studying disease mechanisms, developing protein-based drugs, and refining biotechnology applications. MtoZ Biolabs provides integrate Peptide Structure Determination Service.

Analysis Workflow

Sample Preparation

Extract and purify target peptides using high-performance liquid chromatography (HPLC) or other suitable techniques. Desalt and concentrate purified peptides to meet structural analysis requirements.

Sequence Analysis

(1) Select appropriate enzymes to digest peptide samples based on sequence characteristics, producing fragments for mass spectrometry analysis.

(2) Use HPLC to separate enzymatically cleaved peptides.

(3) Determine peptide fragments' amino acid composition and sequence via Edman degradation or tandem mass spectrometry (MS/MS), revealing the primary peptide structure.

Structural Analysis

Measure the peptides ultraviolet absorption by circular dichroism (CD), identifying their internal secondary structure elements such as -helices and -folds.

Revealing Protein Structure

Peptide structure determination identifies protein structures, including secondary, tertiary, and quaternary structures.

Studying Protein Function

Since protein structure closely correlates with function, peptide structure determination illuminates these relationships.

New Drug Development

By identifying drug-target protein interactions, peptide structure determination provides crucial insights for drug design.

Disease Diagnosis and Treatment

Peptide structure determination helps identify disease-specific biomarkers, enhancing early diagnosis and treatment efficacy.

Deliverables

In the technical report, MtoZ Biolabs will provide you with a detailed technical information, including:

Experimental ProceduresRelevant Mass Spectrometry ParametersPeptide Structure Determination Detailed InformationMass Spectrometry ImagesRaw Data

The immunopeptidome comprises all short peptides presented on the cell surface by HLA-I and II molecules for T-cell recognition. Immunopeptidomics analysis aims to explore the dynamics and composition of both type I and II immune peptides. Comprehensive characterization of the immunopeptidome aids in developing new therapies for cancer, immune diseases, and infectious diseases. MtoZ Biolabs provides integrate Immunopeptidomics Service.

Immune peptides are the complete set of peptides presented by nucleated cells through type I and II proteins within the HLA or MHC antigen presentation pathways. They are essential in defining immunogenic epitopes during immune responses. Characterizing these immune peptides can help identify targets for personalized cancer immunotherapies, including neoantigens arising from tumor-specific mutations or tumor-associated antigens. Moreover, this knowledge facilitates the development of novel mRNA and peptide vaccines and cell-based therapies.

MtoZ Biolabs offers a comprehensive high-resolution mass spectrometry-based immunopeptidomics analysis and neoantigen discovery solution, including proprietary, highly sensitive immune peptide enrichment and identification protocols. Our approach helps identify over 10, 000 type I and over 10, 000 type II peptides. Using our optimized high-throughput platform, immunopeptidomics analysis enables reliable identification and quantification from minimal samples. This service supports large-scale studies, empowering researchers to uncover solutions for cancer, immune diseases, and infectious diseases and identify unexplored therapeutic targets.

Services at MtoZ Biolabs

MtoZ Biolabs' comprehensive neoantigen discovery solution offers peptide synthesis, functional screening of neoantigen candidates for personalized cancer vaccines and T-cell therapies, animal testing of personalized cancer vaccines, and clinical-stage vaccine immunogenicity trials.

Deliverables

A Comprehensive List of All Identified Immune Peptides, Neoantigen Peptides, and Tumor-Associated Antigen PeptidesImmunopeptidome Data Quality Analysis Reports, as Presented in Case Studies

Sample Submission Requirements

Biomarkers are measurable traits that serve as indicators of normal biological activities, disease processes, or pharmacological responses to therapeutic interventions. Peptides are particularly suitable as biomarkers due to their ability to circulate among various body compartments. Many disease processes can be inferred from the characteristic peptide profiles and pathological changes observed in different body fluids. Moreover, variations in peptide abundance associated with various diseases are detectable. MtoZ Biolabs provides integrate Peptide Biomarker Discovery & Validation Service.

Two prevalent approaches to developing peptide biomarkers are pattern recognition and single/oligo biomarker assay. Pattern recognition does not require identifying the candidate biomarkers and relies solely on mass spectrometry's exceptional specificity and sensitivity to derive patient information. In contrast, the single/oligo biomarker assay is more effective as their expression levels can be verified across various biological samples. Mass spectrometry-based methods can accurately determine analyte concentrations and feature precise mass measurement and sequencing capabilities.

Figure 1. Peptide Biomarkers Analysis

To screen and validate peptide biomarkers, it is necessary to identify the peptides in a sample (qualitative analysis) and quantify their concentrations (quantitative analysis). Effective screening requires integrating both steps. Numerous analytical techniques are employed in biomarker detection, among which the combination of mass spectrometry and liquid chromatography (e.g., UPLC and nanoLC) stands out for its high resolution, sensitivity, throughput, and quantification capabilities.

MtoZ Biolabs employs the Thermo Fishers Orbitrap Fusion Lumos mass spectrometry system along with nanoLC chromatography for advanced peptide biomarkers analysis services.

Peptides, structurally akin to proteins, are molecules composed of amino acid sequences. They play pivotal roles in regulating blood pressure, modulating pain perception, and managing glucose metabolism, yet are significantly smaller than proteins. Endogenous peptides typically arise from the cleavage of precursor proteins by various enzymes. Research into the collective peptide expression under specific physiological conditions is termed peptidomics. MtoZ Biolabs provides integrate Mass Spectrometry-Based Peptide Identification Service.

Peptide identification aims to achieve "Peptide-Spectrum Matches" by analyzing peptide sequence information to ascertain sequences or post-translational modifications. Tandem mass spectrometry, extensively employed in peptide identification, combines with bioinformatics for peptide sequencing in proteomics. This involves matching experimental spectral data against theoretical data from protein databases. A current focus of research is matching proteins with unknown sequences to database entries.

Analysis Workflow

Services at MtoZ Biolabs

Peptide Extraction from Test MaterialsChromatographic Separation of PeptidesPrimary Mass Spectrometric Analysis of Ionized Peptides Mass-to-Charge RatiosSecondary Mass Spectrometric Analysis of Fragmented Peptide IonsBioinformatics Analysis

Gel sealing in plastic film or gel strips in sealed tubes. Maintain moisture in sealed samples without excessive water or buffer.

Considerations

Mandatory Glove Use for Gel CuttingCleanliness of Cutting BladesPrecision in Cutting Gel Strips to Minimize Background NoiseStringent Avoidance of Keratin Contamination from Handling and Environmental Exposure

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

Experimental ProceduresMass Spectrometry ParametersMass Spectrometry ImagesRaw DataDetailed Information of Identified Proteins

Peptide purity is defined as the ratio of the target peptide to all analytes, essentially reflecting the proportion of the target peptide to impurities. It is typically determined using HPLC analysis at 220nm, the peak absorption wavelength for peptide bonds. The purity of peptides critically influences research outcomes, with potential contaminants arising during chemical synthesis, culturing, or the extraction and purification phases. Impurities may include peptides with incomplete sequences, truncated sequences, incompletely deprotected peptides, and by-products from synthesis or final cleavage processes. MtoZ Biolabs provides integrate Peptide Purity Analysis Service.

Applications

Services at MtoZ Biolabs

RP-HPLC

Chromatograms from reverse phase HPLC are utilized to assess peptide purity by quantifying the peak area of the fully purified peptide relative to the total detected peak areas, facilitating the determination of by-product quantities and proportions.

Mass Spectrometry

Chromatograms from RP-HPLC are utilized to assess peptide purity by quantifying the peak area of the fully purified peptide relative to the total detected peak areas, facilitating the determination of by-product quantities and proportions.

Amino Acid Analysis (AAA)

Determines the amino acid composition in peptide or protein products, ensuring experimental accuracy and consistency.

MtoZ Biolabs delivers peptide purity analysis services leveraging RP-HPLC and mass spectrometry, dedicated to high-quality results. Each project requires a unique approach, so please engage our technical team to discuss specific needs.

The objective of peptidomics analysis extends beyond identifying and verifying all endogenous peptides in the biological samples under study. It also involves comparing the expression levels of target peptides across specific biochemical processes. Mass spectrometry facilitates a comprehensive analysis of these peptides. MtoZ Biolabs provides integrate Peptidomics Service.

Peptidomics and proteomics have similar strategic approaches in research; however, notable differences exist. In peptidomics, to accurately identify native peptides, including those with post-translational modifications, specific digestive enzymes are generally avoided. This enables direct identification through primary MS and secondary MS.

Advancements in MS/MS sequencing have increasingly supplanted the traditional Edman degradation technique for peptide identification. Mass spectrometry has become pivotal for identifying peptides in various tissue extracts, including bioactive peptides. Size exclusion chromatography (SEC) is employed to remove most proteins from biological samples, body fluids, and tissue extracts, thereby enhancing the precision and repeatability of LC-MS in peptide identification. MtoZ Biolabs offers tailored peptidomics analysis services to meet specific customer requirements.

In cell biology and biochemistry, post-translational modifications (PTMs) critically influence the stability, activity, localization, and functions of proteins. Among these, lactylation stands out as a significant PTM. This modification can occur on any protein residue with a free amino group, such as lysine, arginine, and tyrosine. During this process, lactate reacts with these amino acids to form an ester linkage, thereby altering protein structure and function. This change impacts several biological processes, including cell signaling, gene expression, and metabolism. Accurate identification and characterization of lactylation sites are essential for understanding their biological implications and for the development of related applications.MtoZ Biolabs provides integrate Protein Lactylation Modification Analysis Service.

Mass spectrometry is crucial for identifying lactylated proteins and studying their properties. This technique allows for precise determination of lactylation sites and explores the modification's impact on protein structure and function. Additionally, mass spectrometry is also used to screen for new lactylation sites, providing deeper insights into protein lactylation studies. Moreover, research on lactylation modification has a broad application value, including but not limited to understanding the structure and function of proteins, studying the pathogenesis of diseases, and the development of novel drugs, and helping us understand the complexity and dynamics of biological systems.

Figure 1. Mass Spectrometric Analysis of Protein Lactylation Modification

At MtoZ Biolabs, using the latest Thermo's Obitrap Exploris 240 mass spectrometer in combination with Nano-LC technology, we have developed a precise analytical platform for protein lactylation. This platform is adept at pinpointing lactylation sites and assessing their dynamic changes under various conditions, thereby revealing their effects on protein structure and function. Whether you aim to investigate the fundamental mechanisms of lactylation or its role in disease progression, MtoZ Biolabs offers comprehensive, efficient, and tailored analytical solutions. Contact us for more information on our protein lactylation modification analysis services.

Analysis Workflow

Protein Sample Preparation

Extract proteins from biological samples.

Protein Digestion

Digest proteins into smaller peptides using enzymes like trypsin or chymotrypsin.

Liquid Chromatography Separation

Further separate enriched peptides via high-performance liquid chromatography.

Peptide Enrichment

Enrich lactylated peptides using techniques like antibody enrichment or specific solid phase extraction.

Mass Spectrometric Analysis

Determine peptide masses and their fragmentation patterns to identify amino acid sequences, lactylation sites, and obtain quantitative data.

Angel, T. E. et. al. Chem. Soc. Rev. 2012.

Figure 2. Lactylation Modification Analysis Process

Service Advantages

Sensitivity and Accuracy

Detect lactylation modifications with high precision even at low abundances.

Localization of Modification Sites

Identify lactylation on proteins and precisely determine the affected amino acid residues.

Quantitative Capabilities

Assess the relative or absolute abundance of lactylation modifications to understand their biological impacts.

High-throughput Capability

Identify and quantify numerous lactylation sites in a single experiment.

Compatibility

Combine with other techniques like immunoprecipitation and bioinformatics for more comprehensive analyses.

Deliverables

Protein deamidation involves the conversion of amide groups (-CONH2) in amino acids like glutamic acid and aspartic acid into their corresponding acids, -ketoglutarate and glutamic acid, by losing an ammonia molecule (NH3). This non-enzymatic post-translational modification is crucial for understanding proteins' charge distributions, functions, stability, and interactions with other molecules, as well as their roles in diseases. For example, -amyloid proteins associated with Alzheimer's disease may become more prone to forming fibrils post-deamidation. Advances in molecular biology and mass spectrometry now allow for detailed analysis of such modifications. Using mass spectrometry, researchers can pinpoint exactly where deamidation occurs within protein sequences, which is essential for exploring protein functions, structures, and interactions. MtoZ Biolabs provides integrate Protein Deamidation Modification Analysis Service.

Figure 1. Protein Deamidation Modification Process

Protein deamidation analysis via mass spectrometry is a sophisticated approach in protein research. This method involves inducing deamidation through specific enzymes or chemicals, converting aspartic and glutamic acids' carboxyl groups into amine groups. This change reduces the mass of these amino acids by about 1Da, creating distinct mass-to-charge ratios detectable in mass spectrometry. This differential allows for the precise identification and quantification of deamidation modifications in proteins. MtoZ Biolabs offers a high-resolution mass spectrometry-based platform for this analysis, ensuring high-quality, comprehensive services for studying cellular signaling, protein interactions, and disease mechanisms.

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

Experimental ProceduresRelevant Experimental ParametersDetailed Information on Protein Deamidation ModificationsMass Spectrometry ImagesRaw Data

Protein phosphorylation is a reversible post-translational modification where amino acid residues in proteins are phosphorylated by kinases, which attach covalently bonded phosphate groups. This modification alters proteins' conformation, potentially activating, deactivating, or changing their functions. Research into protein phosphorylation enhances our understanding of biological processes and helps to define the mechanisms of diseases linked to abnormal phosphorylation levels. Due to the low concentration and broad dynamic range of phosphorylated proteins in biological samples, enrichment of phosphorylated peptides is essential before performing quantitative proteomics to increase the detectability of these modifications. MtoZ Biolabs provides integrate Multipathway Phosphoproteomics Service.

MtoZ Biolabs employs the Multi-Pathway Enrichment Kit from CST company, along with Thermo Fisher's Orbitrap Fusion Lumos mass spectrometer and nanoLC, to offer comprehensive services in multipath protein phosphorylation proteomics. Clients need only specify their research goals and submit their samples; MtoZ Biolabs manages all subsequent steps including protein extraction, digestion, phosphorylated peptide enrichment, peptide separation, mass spectrometric analysis, analysis of raw data, and bioinformatics.

Analysis Workflow

Sample Submission Requirements

For Tissue Samples

Ship on dry ice; minimum requirements are 200 mg for plant tissues, 1 mL for blood (plasma should be anticoagulated with EDTA), 0.5 mL for serum, 2 mL for urine, 1 g for animal tissues, and 5*107 cells for cell samples. Yeast and microorganisms should have a dry weight of 200 mg.

For Protein Samples

Ensure a minimum of 1 mg total protein. Use standard lysis solutions for tissue and cell extraction.

Sample Shipping

Transport samples with adequate dry ice and opt for expedited shipping to minimize degradation risks during transit.

Sample Testing

We evaluate all samples before commencing the actual experiments. Testing must confirm sample integrity before proceeding with formal analyses.

Methylation represents one of the most prevalent post-translational modifications, chiefly affecting transcription factors and histones, though it also modifies a subset of cytoplasmic proteins. This modification includes monomethylation, symmetric/asymmetric dimethylation, and trimethylation on arginine (R) and lysine (K) residues. Arginine methylation plays critical roles in RNA processing, gene transcription, DNA damage repair, protein translocation, and signal transduction, whereas lysine methylation primarily influences histone function and epigenetic regulation of gene transcription. Targeted enrichment of methylated peptides using modification-specific antibodies, followed by LC-MS/MS analysis, facilitates extensive quantitative and qualitative assessment of protein methylation. MtoZ Biolabs provides integrate Quantitative Methylomics Service.

MtoZ Biolabs employs the Orbitrap Fusion Lumos mass spectrometer and nanoLC from Thermo Fisher for comprehensive protein methylation identification services. Clients need only submit their experimental aims and samples; MtoZ Biolabs handles all subsequent steps, including protein extraction, digestion, methylation specific peptide enrichment, peptide separation, mass spectrometric analysis, analysis of raw data, and bioinformatics.

Analysis Workflow

Plant Tissues (roots, xylem, phloem): > 5 g

Animal Tissues: > 200 mg wet weight

Microorganisms: > 2 g wet weight

Body Fluids (saliva, amniotic fluid, CSF): > 10 mL

Serum: > 500 L

Urine: > 50 L

Protein Extracts: > 2 mg/mL concentration, minimum total 1 mg, free of interfering substances like nucleic acids, lipids, and polysaccharides.

For Tissue Samples

Ship on dry ice.

For Protein Samples

Use standard lysis solutions for tissue and cell extraction.

Sample Shipping

Transport samples with adequate dry ice and opt for expedited shipping to minimize degradation risks during transit.

Sample Testing

We evaluate all samples before commencing the actual experiments. Testing must confirm sample integrity before proceeding with formal analyses.

Our team, seasoned in protocol optimization, ensures top-quality results in protein methylation studies. For specific inquiries, please contact us directly.

Service Advantages

High-Throughput

Capability to identify and quantify thousands of proteins simultaneously.

Complete Coverage

Digestion with 2-3 enzymes guarantees comprehensive protein analysis.

Applicability to Low-Abundance Methylated ProteinsState-of-the-Art Equipment

The Orbitrap Fusion Lumos, noted for its superior resolution and sensitivity.

Services at MtoZ Biolabs

Ion ChromatographyHigh Performance Liquid Chromatography (HPLC)Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS)

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

Experimental ProceduresRelevant Mass Spectrometry ParametersDetailed Information on Identified Methylation SitesMass Spectrometry ImagesRaw Data

Protein oxidative modifications are generally induced by reactive oxygen species (ROS) such as superoxide anions, hydrogen peroxide, and hydroxyl radicals. These ROS are generated through multiple pathways in living organisms, including mitochondrial respiration, enzymatic redox reactions, and exposure to radiation. While protein oxidative modification is a natural physiological process, excessive ROS production or insufficient removal can cause excessive oxidation, impairing protein function and potentially leading to cellular damage. MtoZ Biolabs provides integrate Protein Oxidative Modification Analysis Service.

Advancements in molecular biology and biochemistry have positioned proteomics mass spectrometry as the primary method for investigating protein oxidative modifications. This technique allows for the precise identification of oxidatively modified amino acid residues and their chemical structures, facilitating detailed exploration into the types and mechanisms of these modifications. Employing mass spectrometry enables high-throughput screening and quantitative analysis of protein modification sites, offering crucial insights into cellular oxidative stress responses and the mechanisms underlying associated diseases.

Figure 1. Proteomic Oxidative Modification Mass Spectrometric Analysis

MtoZ Biolabs utilizes the Thermos Obitrap Fusion Lumos mass spectrometer in conjunction with Nano-LC technology to perform highly accurate analysis of protein oxidative modifications. This approach supports both quantitative and qualitative assessments, providing detailed information on modification types and locations. We accept a variety of protein samples and provide tailored one-stop solutions to meet diverse research needs. Free project evaluation!

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

Experimental ProceduresRelevant Experimental ParametersDetailed Information on Protein Oxidative ModificationsMass Spectrometry ImagesRaw Data

Small ubiquitin-like modifier (SUMO) is a post-translational modification of proteins that refers to the covalent attachment of a small ubiquitin-associated modifier to a protein. Unlike ubiquitination, which targets proteins for degradation, sumoylation modulates protein function across several cellular processes including nucleocytoplasmic transport, transcription regulation, apoptosis, protein stability, stress response, and cell cycle progression. Despite low amino acid sequence similarity between SUMO and ubiquitin, they share very similar structural folds. SUMOs are about 100 amino acids in length, with variations in sequence length and mass among different family members and organisms. MtoZ Biolabs provides integrate Protein Sumoylation Identification Service.

Sumoylation is pivotal in controlling the subcellular localization of various proteins. Depending on the specific protein, sumoylation can occur in either the cytoplasm or nucleus. For instance, sumoylation can regulate the transport of the ribonucleoprotein RanGAP1 within the nuclear pore complex (NPC). Additionally, sumoylation affects the transcription process, as the activity of several transcription factors depends on their interaction with promyelocytic leukemia (PML) bodies and nuclear bodies (NBs), which require the sumoylation of PML protein for their assembly. Moreover, sumoylation plays a crucial role in chromosome congression and kinetochore assembly. If the function of SUMO-1 is abnormal, it can disrupt the correct distribution of chromatin during replication.

Figure 1. Protein Sumoylation Identification

MtoZ Biolabs leverages the Thermo Fisher's Orbitrap Fusion Lumos mass spectrometry platform, coupled with nanoLC-MS/MS chromatography, to deliver a comprehensive protein sumoylation analysis service. Clients are invited to submit their experimental objectives and samples, and our team will manage all aspects of the project thereafter.

Analysis Workflow

Protein Digestion in Gel or SolutionEnrichment of Sumoylated Proteins Through Antibodies Specific to Sumoylation MotifsHPLC Separation Followed by ESI-TOF MS/MS AnalysisAnalysis of Mass Spectrometry Data

Functional Annotation and Enrichment AnalysisCluster AnalysisNetwork AnalysisStatistical AnalysisAnalysis of Post-Translational Modification in Proteomics

Acylation is a post-translational modification involving the attachment of acyl groups, such as acyl-CoA, to proteins. This modification plays a pivotal role in regulating key biological processes including epigenetics, energy metabolism, protein trafficking, and molecular interactions. Acylation research is a dynamic field in life sciences, with studies expanding beyond acetylation to include propionylation, malonylation, glutarylation, succinylation, and crotonylation. Quantitative proteomics of acylation provides insights into the vital link between protein acylation changes and biological functions, offering valuable information for uncovering life mechanisms, identifying clinical disease markers, and targeting drug discovery. MtoZ Biolabs provides integrate Quantitative Acetylomics Service.

The acylation quantitative proteomics process begins by enzymatically digesting protein samples into peptides. These peptides are then separated using liquid chromatography to simplify the mixture. Specific acylation modifications are enriched using targeted antibodies or kits, followed by quantification with LC-MS/MS.

MtoZ Biolabs employs the Thermo Fishers Orbitrap Fusion Lumos mass spectrometry system alongside nanoLC platform for comprehensive acylation quantitative proteomics services. We invite researchers to share their experimental goals and submit their samples, after which our team manages all aspects of the analysis. This includes protein extraction, digestion, peptide enrichment, chromatographic separation, mass spectrometric analysis, and subsequent bioinformatics evaluations.

Sample Submission Requirements

For Tissue Samples

Ship on dry ice.

For Protein Samples

Use standard lysis solutions for tissue and cell extraction.

Sample Shipping

Transport samples with adequate dry ice and opt for expedited shipping to minimize degradation risks during transit.

Sample Testing

We evaluate all samples before commencing the actual experiments. Testing must confirm sample integrity before proceeding with formal analyses.

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

Experimental ProceduresRelevant Spectrometric ParametersComprehensive Details on Identified Acylation SitesMass Spectrometry ImagesRaw Data

S-nitrosylation (SNO) is the covalent attachment of some nitroso (NO) groups to sulfhydryl residues (S) in proteins, forming S-nitrosothiols (SNOs). These groups are subsets of specific cysteine residues, leading to the formation of S-nitrosylated proteins. SNOs have a short half-life in the cytosol due to various reductases, such as glutathione (GSH) and thioredoxin, which act to remove the nitroso groups. Therefore, SNOs are predominantly stored in protected environments like plasma membranes, vesicles, intercellular substance, and the folds of lipophilic proteins to prevent denitrosylation. For instance, caspases involved in apoptosis are stored as SNOs within the intermembrane spaces of mitochondria. MtoZ Biolabs provides integrate S-Nitrosylation Analysis Service.

SNO process is reversible, with denitrosylation, an enzyme-catalyzed reaction, reversing the modification. SNO is selective, targeting specific cysteine residues. Improper SNO can result in protein misfolding, synaptic damage, and apoptosis. Furthermore, abnormalities in SNO signaling are associated with the development of diseases such as Alzheimer's and cardiovascular conditions.

Given the challenges in detecting S-nitrosylated cysteines due to the instability of SNOs and the presence of multiple cysteines in proteins, MtoZ Biolabs offers an advanced, sensitive analytical platform for analyzing S-nitrosylated cysteines in both eukaryotic and prokaryotic systems. We have also enhanced our detection methods to provide quicker and more sensitive analyses of these modifications.

Analysis Workflow

Protein Hydrolysis in Gel or SolutionEnrichment of S-Nitrosylated Cysteines Using Specific AntibodiesHPLC Separation Followed by ESI-TOF MS/MS AnalysisMass Spectrometry Data Analysis

Ion ChromatographyHigh-Performance Liquid Chromatography (HPLC)Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS)

Top-down mass spectrometry (TDMS) is highly effective for comprehensive analysis of protein post-translational modifications (PTMs) because it evaluates intact proteins or peptides without requiring hydrolysis. This method is particularly valuable in the analysis of biopharmaceutical proteins, such as monoclonal antibodies and recombinant proteins, where it preserves many unstable modifications that are often lost during collision-induced dissociation (CID) cleavage in shotgun approaches. MtoZ Biolabs provides integrate Top-Down PTMs Analysis Service.

Figure 1. Top-Down PTMs Analysis

Employing techniques like electron capture dissociation (ECD) and electron transfer dissociation (ETD), TDMS excels at accurately measuring the masses of whole proteins or peptides, quantifying various forms of modified proteins, creating detailed modification maps with high coverage, identifying unanticipated PTMs, and sequencing multiple modifications. Despite its relative novelty and the technical challenges it faces in sample preparation and processing throughput, top-down proteomics offers distinct benefits that render it crucial for detailed PTMs characterization.

At MtoZ Biolabs, our team of skilled proteomics scientists and technicians leverages extensive experience in top-down proteomics to deliver superior PTMs characterization services.

With ongoing advancements in proteomics mass spectrometry technology, there has been a notable increase in the precision of analytical instruments, resulting in a richer dataset. While this detailed protein data is invaluable for deeper scientific investigation, the vast volumes of data produced by high-throughput proteomic screenings introduce substantial challenges in analyzing protein sample characteristics. Due to the complexity of handling such extensive mass spectrometry data manually, it is imperative to apply advanced bioinformatics techniques for effective analysis. MtoZ Biolabs boasts a team of bioinformatics analysts who excel in the extraction of insights from proteomics data, particularly focusing on the analysis of biological pathways and networks to swiftly identify extensive protein interactions. Leveraging advancements in bioinformatics analytical methods, we have developed a comprehensive platform for proteomics data analysis. Our suite of bioinformatics solutions encompasses quality assessment of omics data, differential expression analysis, annotation and enrichment analysis through GO, KEGG, and COG, as well as protein clustering and the analysis of interaction networks and pathways at multiple levels. Furthermore, we offer tailored bioinformatics services based on client specifications and scholarly references, enabling the extraction of valuable biological insights from high-throughput experimental data. This support aids our clients in areas such as drug development, toxicity testing, and the identification of disease markers. MtoZ Biolabs provides integrate Proteomics Bioinformatic Analysis Service.

Services at MtoZ Biolabs

Proteomic Data QualityDifferential Protein Statistical AnalysisGO Functional Annotation and Enrichment AnalysisKEGG Pathway and Enrichment AnalysisCOG Functional Annotation and Enrichment AnalysisDifferentially Expressed Proteins Custer AnalysisProtein Interaction Analysis

To validate the selection of differentially expressed proteins or characteristic differentially expressed proteins, cluster analysis can be performed to group proteins based on expression trends, facilitating more intuitive proteomic data analysis. MtoZ Biolabs provides integrate Differential Proteins Clustering Analysis Service.

Services at MtoZ Biolabs

Hierarchical Cluster AnalysisK-means Cluster Analysis

The COG (Clusters of Orthologous Groups) database is an early and widely used resource for the homologous classification of gene products, based on extensive comparisons of protein sequences from various organisms. MtoZ Biolabs provides integrate COG Functional Annotation Analysis Service.

Differential protein analysis can significantly enhance the discovery of new biomarkers, improving the accuracy of biomarker identification and providing valuable insights for clinical disease analysis. MtoZ Biolabs offers precise differential protein analysis based on statistical data, identifying significant differences in protein levels under various physiological conditions and tissues. In the statistical analysis of differential expressed proteins, FC 1.3 (or 1/1.3) is used as the threshold. Proteins with FC 1.3 are considered up-regulated, while those with FC 1/1.3 are considered down-regulated. The number of differential expressed proteins is summarized in the table below. MtoZ Biolabs provides integrate Differential Protein Analysis Service.

Table 1. Statistical analysis of differential expressed proteins

The Venn diagram illustrates the shared and unique differential expressed proteins between two groups:

Figure 1. Venn Diagram of differential Expressed Proteins

Volcano Plot

The volcano plot provides a rapid visualization of the differences in protein expression levels between two sample groups and the statistical significance of these differences.

Figure 2. Volcano Plot of Differential Expressed Proteins

Note: Each point in the volcano plot represents a protein. The x-axis shows the log fold change in expression levels between the two samples, while the y-axis indicates the t-test p-value. A larger absolute value on the x-axis signifies a greater fold change in expression levels between the samples; a higher value on the y-axis denotes more significant differential expression, enhancing the reliability of identified differential expressed genes. Green dots represent down-regulated proteins, red dots represent up-regulated proteins, and black dots indicate non-differential expressed proteins.

The Gene Ontology (GO) is a database created by the Gene Ontology Consortium to provide a standardized vocabulary for gene and protein functions across species, which evolves with ongoing research. It uses a dynamic controlled vocabulary to describe the roles of genes and proteins within cells, offering a comprehensive description of gene and gene product attributes. The GO database is divided into three main categories: biological process (BP), cellular component (CC), and molecular function (MF), each describing potential molecular functions, cellular locations, and biological processes of gene products. Each node in the GO database, identified by unique names such as Cell, Fibroblast Growth Factor Receptor Binding, or Signal Transduction, and a unique identifier like GO:nnnnnnn, is annotated based on protein IDs mapped from the Uniprot database. The proteins are then functionally classified. The number of proteins associated with each GO node in BP, CC, and MF is listed, and statistical graphs of the secondary classification of expressed proteins are provided. MtoZ Biolabs provides integrate GO Functional Annotation and Enrichment Analysis Service.

Services at MtoZ Biolabs

GO Secondary Classification AnalysisGO Levels Classification AnalysistopGO Protein Enrichment Analysis

KEGG (Kyoto Encyclopedia of Genes and Genomes) is a database used for systematic analysis of metabolic pathways and functions of gene products in cells. KEGG combines gene with its expression information as a whole network. KEGG records data from genome, chemical molecule, and biochemical system, including metabolic pathways (PATHWAY), drugs (DRUG), diseases (DISEASE), gene sequences (GENES), and genomes (GENOME). MtoZ Biolabs provides integrate KEGG Pathway Annotation and Enrichment Analysis Service.

Statistics of KEGG Mapping Results for All Identified Proteins

Statistics of KEGG Annotation Results for All Identified Proteins

KEGG Pathway Annotation

In organism, different gene products coordinate to exercise biological functions. Pathway annotation analysis of differentially expressed genes helps to further understand the functions of genes. Pathway annotation diagram for differentially expressed proteins is as follows:

Figure 1. Pathway Diagram of KEGG Annotation Results

Note: Compared to the control group, enzymes marked in red are related to upregulated proteins; enzymes marked in green are related to downregulated proteins; enzymes marked in blue are related to both upregulated and downregulated proteins. The numbers inside the boxes represent the number of enzyme (EC number), and the entire pathway consists of complex biochemical reactions catalyzed by various enzymes. This pathway diagram marks enzymes related to differentially expressed genes in different colors, which focuses on the study of the differential expression of certain metabolic pathway-related proteins according to the difference among subjects and explains the root causes of phenotypic differences through the pathway.

KEGG Pathway Classification

The annotation results of differentially expressed genes in KEGG are classified according to the pathway types in KEGG, results are shown in the following figure:

Figure 2. KEGG Classification Diagram of Differentially Expressed Proteins

Note: The vertical axis is the name of the KEGG metabolic pathway, and the horizontal axis represents the number of proteins annotated to that pathway and their proportion of the total annotated proteins.

KEGG Pathway Enrichment

Analysis of whether differentially expressed proteins are over-represented in a certain pathway is termed pathway enrichment analysis for differentially expressed proteins. We use Kobas software to perform KEGG pathway enrichment analysis for differentially expressed proteins. The results of the KEGG pathway enrichment analysis for differentially expressed proteins are shown in the following figure:

Figure 3. KEGG Pathway Enrichment Statistics Diagram of Differentially Expressed Proteins

Note: Each point in the diagram represents a KEGG pathway, with the pathway name on the left axis. The horizontal axis represents the enrichment factor, a ratio that the proportion of differentially expressed proteins annotated to the pathway to the proportion of proteins in that species annotated to a pathway. The higher the enrichment factor, the more reliable the significance of enrichment of differentially expressed proteins in that pathway.

STRING is a database that records predicted and experimentally validated protein-protein interactions (PPIs) across multiple species, including direct physical interactions and indirect functional associations. By integrating the results of differential expression analysis and the interaction pairs recorded in the database, a differential expression protein interaction network is constructed. During the analysis, differentially expressed proteins are mapped to the STRING database to obtain information on their interaction relationships. As the STRING database includes experimental data, results from text mining of PubMed abstracts, and aggregated data from other databases, as well as predictions made using bioinformatics methods, we select interaction pairs with a combined score greater than 0.4 (Medium) from the search results and use appropriate bioinformatics analysis software to visualize the interaction results. The results are shown in the below. MtoZ Biolabs provides integrate Protein Interaction Network Analysis Service.

Figure 1. Evidence of Differential Expressed Protein Interactions from Different Sources (Evidence)

Figure 2. Graph of Differential Expressed Protein Interaction Strengths (Confidence)

Note: Circles (nodes) represent proteins; Different colors indicate different proteins, and the circles contain the three-dimensional structure of the proteins. Lines indicate the interactions between proteins. Thicker lines indicate stronger interaction relationships. Dark lines indicate relationships in the evidence view circle but no relationships in the actions diagram.

Figure 3: Different Action Modes between Differential Expressed Proteins (Actions)

Proteomics mass spectrometry data is fundamental to proteomics analysis. The quality of the mass spectra is closely related to the accuracy of the retrieval and analysis results. Therefore, the quality assessment of proteomics data is particularly important. To ensure the reliability of bioinformatics analysis, MtoZ Biolabs evaluates proteomic data identified from the following four aspects. MtoZ Biolabs provides integrate Proteomics Data Quality Assessment Service.

Peptide Match Error DistributionPeptide Count DistributionPeptide Length DistributionProtein Molecular Weight DistributionAnalysis of Peptide Match Error Distribution

The error distribution between the molecular weight of all matched peptides and their theoretical molecular weight is shown in the following diagram:

Analysis of Peptide Count Distribution

The distribution of peptide counts in the identified proteins is shown in the following diagram:

Note: The horizontal axis represents the count of peptides, and the vertical axis represents the number of proteins.

Analysis of Peptide Length Distribution

The distribution chart of the lengths of the identified peptides is as follows:

Note: The horizontal axis represents the length of peptides, and the vertical axis represents the number of peptides.

Analysis of Protein Molecular Weight Distribution

Note: The horizontal axis represents the molecular weight of proteins, and the vertical axis represents the number of proteins.

Two-dimensional difference gel electrophoresis (2D-DIGE) is a novel quantitative proteomics technique that evolved from traditional two-dimensional gel electrophoresis (2-DE). The principle of 2D-DIGE for separating mixed proteins is the same as traditional 2-DE. It uses differences in protein isoelectric points and molecular weights to separate protein mixtures. At the same time, the sensitive fluorescent dyes and the internal standard make it significantly better than traditional 2-DE in quantitative proteomics. The fluorescent dyes used in DIGE are Cy2, Cy3, and Cy5, which react with the lysine side chain amino groups of proteins to label the proteins without affecting their isoelectric points and molecular weights. After mixing equal amounts of the labeled proteins, two-dimensional electrophoresis is conducted. The internal standard Cy2 is used to match different gels and eliminate gel viriation, while changes in protein expression levels are reflected by fluorescence intensity of Cy3 and Cy5. As a classic method of quantitative proteomics, 2D-DIGE is widely applied and suitable for various samples. MtoZ Biolabs provides integrate 2D-DIGE Based Protein Quantitative Service.

Figure 1. 2D-DIGE Quantitative Proteomics

MtoZ Biolabs provides SDS-PAGE and 2D-DIGE services. Combining Thermo Fisher's Orbitrap Fusion Lumos mass spectrometer platform and nanoLC-MS/MS nano-liquid chromatography, We provide comprehensive proteomics identification and quantification services for researchers.

Sample Submission Requirements

Both Liquid and Solid Samples are Acceptable

Applications

Suitable for Various Samples

Service at MtoZ Biolabs

Experimental ProceduresRelevant Experimental ParametersGel and Mass Spectrometry ImagesRaw DataResults of the 2D-DIGE Quantitative Proteomics Analysis

Relative quantification of proteins is a MS-based technique that compares the relative proteins expression levels between different samples, analyzing differential protein expression under different conditions and revealing the role of proteins in biological processes. Commonly used methods include stable isotope labeling by amino acids in cell culture (SILAC), isobaric tags for relative and absolute quantification (iTRAQ), and tandem mass tags (TMT). They achieve relative quantification by enzymatic digestion, isotopic labeling, separation, and mass spectrometric analysis. MtoZ Biolabs provides integrate Relative Protein Quantitative Service, MS Based.

MtoZ Biolabs offers protein relative quantification services based on Thermo Fisher's Orbitrap Exploris 240 mass spectrometry platform combined with nanoLC-MS/MS nanoscale chromatography, using multiple relative quantification techniques such as isotopic labeling (e.g., SILAC, TMT, iTRAQ) and label-free methods (e.g., SWATH-MS, DDA). We precisely compare protein expression levels across different experimental conditions or biological samples, identify protein-level changes, and help clients rapidly and accurately assess differences in protein expression levels to reveal the molecular mechanisms of biological processes, disease onset, and drug therapy.

Analysis Workflow

Sample Preparation

Proteins extraction from different biological samples using suitable methods according to project requirements, followed by purification, concentration, and quality assessment to ensure the sample meets experimental requirements.

Protein Digestion

Protein digestion by various enzymes into peptide for subsequent mass spectrometry analysis.

Isotopic Labeling

Peptide labeling by isotopic labeling methods such as SILAC, iTRAQ, or TMT.

Peptide Separation

Peptide separation by HPLC (e.g., RP-HPLC).

Mass Spectrometric Analysis

Analysis and detection of peptides by high-resolution mass spectrometry to obtain data on mass, m/z and other information.

Data Analysis

Analysis of mass spectrometry data by specific bioinformatics software to identify and relatively quantify proteins to find significantly different proteins by comparing differential protein expression under different conditions.

Service Advantages

High-Throughput Data Acquisition

Advanced Orbitrap Exploris 240 mass spectrometer with high-sensitivity, high-resolution protein detection to quickly acquire large amounts of protein data.

Accurate Quantitative Analysis

Efficient data processing workflows and software for accurate and reliable protein quantification.

High Sensitivity

Ability to detect low-abundance proteins with high sensitivity.

Broad Applicability

Suit for a wide range of biological samples, including cells, tissues, and body fluids to study various biological processes.

Customized Analytical Solutions

One-stop services including experimental design, sample processing and data analysis according to project requirements.

Applications

Gene Expression Regulation Studies

Compare protein expression differences under different conditions to study the effects of post-transcriptional regulation and post-translational modifications on gene expression.

Signal Transduction Pathway Studies

Use quantitative analysis of differential protein expression of signal molecules to reveal activation and inhibition mechanisms of signaling pathways.

Comparative Proteomics Studies

Compare protein expression differences among species or subtypes to explore genetic variation, evolutionary relationships, and functional differences in species.

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Data-independent acquisition (DIA) is a non-dependent data scanning mode, a holographic mass spectrometry data acquisition mode based on the electrostatic field orbitrap. After primary mass spectrometric detection, all ionized compounds of a given sample that fall within a specified mass range are fragmented in a systematic and unbiased fashion. The fragment ions of all precursor ions are collected, and quickly scaned within precursor isolation window to perform protein qualitative and quantitative analysis. Compared to data-dependent acquisition mode (DDA), DIA has better accuracy and reproducibility. The SWATH technique is a DIA technique that integrates the high-throughput detection of shotgun proteomics with the precise quantitative analysis of parallel reaction monitoring (PRM). It can quantify almost all detectable molecules in complex samples. MtoZ Biolabs provides integrate DIA based Protein Quantitative Service.

MtoZ Biolabs offers DIA quantitative proteomics service, including the AB SCIEX Triple-TOF 5600 plus high-resolution mass spectrometry system, which features fast scanning speed and quantitative sensitivity of tandem quadrupole mass spectrometry systems and integrates high resolution, accurate mass stability, high sensitivity, and high-speed scanning of the mass spectrometry system. You only need to send us your samples, and we will take care of all subsequent project matters, including protein extraction, protein digestion, peptide separation, mass spectrometric analysis, raw mass spectrometric data analysis, and bioinformatics analysis.

Applications

It can be applied to various bioscience research in agriculture, forestry, environment, food, and medicine to build biological sample database and discover biomarkers.

Service at MtoZ Biolabs

Experimental ProceduresRelevant Mass Spectrometric ParametersMass Spectrometric ImagesRaw DataProtein Differential AnalysisBioinformatics Analysis

In scientific research, there are two main types of analysis: qualitative and quantitative analysis. Between qualitative and quantitative analysis, there is a semi-quantitative analysis, which yields approximate measurement value rather than precise one. Semi-quantitative analysis is often used when direct measurement of results is challenging while the conclusions drawn from inference are insufficient, especially when quantitative data are likely to fluctuate periodically. MtoZ Biolabs provides integrate Semi Quantitative Proteomic Analysis Service.

Services at MtoZ Biolabs

GO Function Annotation and Enrichment AnalysisCOG Function Annotation and Enrichment AnalysisDifferential Protein Expression Clustering AnalysisProtein-protein Interaction Network AnalysisDifferential Protein Statistics Analysis

Proteomics is a scientific field that studies the composition, structure, function, and interactions of all proteins in an organism. Its goal is to establish a comprehensive protein information library and understand their roles in life. However, the proteins whose biological functions have been thoroughly studied are still few, and some protein-coding genes still lack relevant annotations. Nonsynonymous single nucleotide polymorphisms, a large variety of proteins produced by alternative splicing and post-translational modifications of proteins, make gene annotation more difficult. The emergence of high-throughput gene knockout technology provides a powerful tool for the detection of large-scale protein and their forms. High-throughput gene knockout technology can be used to silence or knockout specific genes to observe changes in the proteome. Combined with high-resolution mass spectrometry, it can be used to analyze protein composition, modifications, and relative abundance. MtoZ Biolabs provides integrate High-Throughout Proteomics Analysis Service for Gene Knockout.

The combination of high-throughput gene knockout technology and proteomics provides a new perspective for biological research. By combining these two technologies, researchers can study the impact of specific gene knockouts on the proteome, and identify proteins associated with the knocked-out genes, which help better understand the interactions between genes and proteins, and reveal protein regulatory networks. For example, researchers can determine how the knockout of a specific gene affects protein expression in cancer cells to discover new cancer treatment methods. Furthermore, this combination can also be used to study the mechanisms of various diseases such as neurodegenerative disease, immune disease, and metabolic disorder.

MtoZ Biolabs, based on the high-resolution proteomics analysis platform combined with high-throughput gene knockout technology, can provide you with a comprehensive solution from gene knockout to proteomics analysis. Our advanced CRISPR/Cas9 system can achieve single/multiple gene knockouts of human/mouse cell lines, primary cells, immune cells, and iPS cells, frameshift mutations, and large sequence deletions. When KO cells obtained, they are cultured and the proteins expressed by them are extracted. Using a high-resolution mass spectrometry platform for their proteomics analysis, the specific role of the knocked-out gene at the protein level is elucidated, revealing the mechanism how genes regulate proteins. MtoZ Biolabs aims to provide you with the highest quality scientific research service. We look forward to cooperating with you. Welcome to contact to learn more about our service.

Service Advantages

High Knockout Efficiency

Optimized CRISPR/Cas9 system with over 80% target knockout efficiency at 70%; after gene knockout, we use Sanger sequencing and in-depth proteomics technology to doubly verify the knockout efficiency to ensure the accuracy of the data.

Comprehensive Proteomics Analysis Platform

Our proteomics analysis platform is equipped with comprehensive sample pre-treatment instruments, high-specificity protein modification enrichment platform, ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry, and protein N-terminal sequence analyzer, which can achieve efficient protein extraction at the omics level, high-efficiency enrichment of specific modified peptides, and protein identification, relative quantification (3D & 4D), and absolute.

Proteomics analysis includes analyzing the structure and function of proteins, post-translational modifications (PTMs), protein localization, protein expression, and the interactions among proteins, etc. Based on different analytical content, different proteomics analysis techniques and strategies can be adopted. MtoZ Biolabs provides integrate Proteomics Analysis Strategy Service.

Mass spectrometry (MS) is one of the most commonly used techniques in proteomics analysis. Currently, proteomics analysis strategies based on MS mainly include bottom-up proteomics and top-down proteomics. In bottom-up proteomics, proteins need to be digested into peptides before mass spectrometric analysis, and this strategy is adopted by common label-free and labeling quantitative proteomics techniques. In top-down proteomics, protein are not digested but are directly analyzed, which can preserve most of the features of proteins with unstable structures that are mostly destroyed in the bottom-up strategy, and also reduces the time consumption of digestion. Researchers can choose the appropriate strategy for proteomics analysis according to different research needs.

MtoZ Biolabs provides various proteomics analysis services using the Thermo Fisher Orbitrap Fusion Lumos mass spectrometry platform combined with nanoLC-MS/MS nanoscale chromatography.

Services at MtoZ Biolabs

Top-down ProteomicsDIA-PRM ProteomicsPCT-DIA Proteomics

Data-independent acquisition (DIA) is a holographic mass spectrometry data acquisition mode based on the electrostatic field Orbitrap and is a data-independent scanning mode. Compared to data-dependent acquisition mode (DDA), DIA technology divides the entire scanning range of mass spectrometry into several windows, selecting, fragmenting, and detecting all ions within each window, providing better accuracy and repeatability. PRM (parallel reaction monitoring) is an ion monitoring technology based on high-resolution and high-precision mass spectrometry, which is currently the mainstream method for targeted proteomics data acquisition. By selectively detecting specific or target peptides (such as post-translational modification peptides), it enables targeted relative or absolute quantification of target proteins/modification peptides. Combining DIA and PRM for proteomics research can achieve detection of large or complex samples and precise quantification of target proteins, especially improving the detection rate of low-abundance target proteins. MtoZ Biolabs provides integrate DIA-PRM Proteomics Service.

MtoZ Biolabs uses Thermo's latest Obitrap Fusion Lumos mass spectrometer combined with Nano-LC nanoliter chromatography technology to provide a DIA+PRM proteomics analysis service. You only need to send us your samples, and we will handle all subsequent project matters, including protein extraction, protease digestion, peptide separation, mass spectrometric analysis, mass spectrometric raw data analysis, and bioinformatics analysis.

Applications

It can be applied to the fields of agriculture and forestry, environment, food, and medicine to construct biological sample information databases and discover biomarkers in various biological science studies.

Deliverables

Experimental ProceduresMass Spectrometry ParametersMass Spectrometry ImagesRaw DataProtein Differential Level AnalysisBioinformatics Analysis

Pressure cycling technology (PCT) is an efficient biological sample preparation technology and a patented technology developed by Pressure BioSciences Inc (PBI), USA. It uses pressure cycling between atmospheric pressure and ultra-high (liquid) pressure (35, 000 PSI or higher), repeatedly breaking cells and enabling the extraction of important biomolecules such as proteins from various samples (cells and tissues from humans, animals, plants, and microorganisms). Data-independent acquisition (DIA) is a holographic mass spectrometry data acquisition mode based on the electrostatic field Orbitrap and is a data-independent scanning mode. Compared to data-dependent acquisition mode (DDA), DIA technology divides the entire scanning range of mass spectrometry into several windows, selecting, fragmenting, and detecting all ions within each window, providing better accuracy and repeatability. The combination of PCT and DIA technologies allows the study of the proteome of paraffin-embedded or formalin-fixed tissue samples and trace samples. MtoZ Biolabs provides integrate PCT-DIA Proteomics Service.

MtoZ Biolabs, using Thermo's latest Obitrap Fusion Lumos mass spectrometer combined with Nano-LC nanoliter chromatography technology, offers a PCT+PRM proteomics analysis service. You only need to send us your samples, and we will take care of all subsequent project procedures, including protein extraction, protease digestion, peptide separation, mass spectrometry analysis, raw data analysis, and bioinformatics analysis.

Analysis Workflow

Guo, T. et al. iScience. 2019.

Figure 1. PCT+DIA Proteomics

Deliverables

Experimental ProceduresRelevant Mass Spectrometry ParametersMass Spectrometry ImagesRaw DataProtein Differential Level AnalysisBioinformatics Analysis

Plant Proteomics is a branch of proteomics aimed at studying the composition, structure, function, interactions, and regulatory mechanisms of plant proteins. Its research methods are similar to those in proteomics with core techniques used in protein separation, purification, identification, functional annotation, interaction studies, and expression regulation studies. Research in plant proteomics can not only provide a material basis for understanding the law of plant growth, development, and stress adaptation but also offer theoretical foundation and solution for improving crop resistance and quality. By comparing the proteome of plants growing in different conditions, different plant varieties, and individuals under normal and stressed conditions, we can identify specific protein that can serve as targets for genetic improvement and biotechnological strategies or provide molecular markers for the impact of environmental changes on plant growth. MtoZ Biolabs provides integrate Plant Proteomics Service.

Services at MtoZ Biolabs

Protein Identification and Functional Annotation

Using high-resolution mass spectrometry (Thermo Fisher's Q Exactive and Orbitrap mass spectrometers) combined with bioinformatics databases and software, we identify detected proteins and provide detailed functional annotation.

Quantitative Proteomics Analysis

We offer labeled (iTRAQ, TMT, SILAC) and label-free methods for quantitative protein analysis.

Protein-Protein Interaction Studies

Techniques like immunoprecipitation (IP), Co-immunoprecipitation (Co-IP), and affinity purification mass spectrometry (AP-MS) are used to study the interactions between proteins of interest and other proteins. Subsequently, LC-MS/MS is used to identify proteins/protein mixtures in purified samples such as IP, Co-IP, and GST fusion protein pull-down samples.

Protein Post-Translational Modifications (PTMs) Analysis

We offer identification and quantification of PTMs like phosphorylation, glycosylation, ubiquitination, acetylation, methylation, disulfide bonds, nitrosylation, and modification sites.

Differential Protein Expression Analysis

By comparing proteomic data of plants under different treatment conditions, we perform statistical analysis of differential protein expression (Venn diagrams, volcano plots) and clustering analysis (hierarchical clustering analysis, K-means clustering analysis).

Applications

Gene Research

Analyzing differential expression of plant proteins provides information in gene function prediction, gene regulatory network research, and plant physiological ecology.

Agricultural Improvement

Studying protein components and metabolic pathways of plant products can offer references for agricultural biotechnology.

Ecological Research

Understanding the biological mechanisms behind plant adaptive changes can aid ecological protection and biodiversity research.

Plant Disease Diagnosis

Researching the molecular mechanisms of diseases can provide important information for disease diagnosis and prevention.

Sample Submission Requirements

Fresh Tissues: Leaves, roots, stems, flowers, seeds, etc.; at least 100 mgDried Tissues: Plant tissues processed by freeze-drying or drying; at least 50 mgCells: Plant cell suspension or cell lysate; at least 1x107 cellsProtein Extracts: Protein samples extracted, at least 100 g with their concentration not less than 1 g/L

Deliverables

Experimental ProceduresRelevant Mass Spectrometry ParametersDetailed Information on Plant Proteomics AnalysisMass Spectrometry ImagesRaw Data

Cellular proteomics is a scientific discipline that explores the expression, modification, interaction, and function of all proteins within cells. As a significant branch of proteomics, the development of cellular proteomics is closely linked to the progress of proteomics itself. In 1995, the term "proteomics" was introduced, marking the official start of proteomics research. With advancements in mass spectrometry technology, studies on intracellular proteins became increasingly sophisticated. In the early 21st century, cellular proteomics emerged as a subfield of proteomics, gaining widespread attention. Since then, new technologies and methods have continuously driven progress in this field, providing critical insights into cell biology, disease mechanisms, and drug development. MtoZ Biolabs provides integrate Cellular Proteomics Service.

Currently, the primary technical methods in cellular proteomics include: 2D gel electrophoresis and liquid chromatography to separate proteomes into different subgroups for easy identification and quantification; mass spectrometry analysis to identifies protein sequences and modification states using high-resolution tandem mass spectrometry; immunoprecipitation and affinity purification to capture the target proteins and highly purify the proteins using specific antibodies or ligands; and bioinformatics analysis to reveal the biological significance of the proteins by integrating, mining, and visualizing the histological data.

Analysis Workflow

Protein Extraction

Disrupt cells and remove non-protein components such as lipids, nucleic acids, and small metabolites.

Protein Separation and Purification

Use 2D gel electrophoresis and liquid chromatography to separate and purify protein samples

Protein Identification

Apply high-resolution tandem mass spectrometry (MALDI-TOF MS, ESI-MS/MS, etc.) to analyze the purified proteins, identifying their sequence and structural information.

Protein Quantification

Conduct quantitative analysis using isotope labeling methods (e.g., iTRAQ, TMT, SILAC) and label-free methods (e.g., Label Free, SWATH-MS) to compare protein expression differences under different conditions.

Data Processing and Analysis

Process, analyze, and interpret mass spectrometry data using bioinformatics methods, including protein identification, quantitative analysis, protein function annotation, interaction network analysis, and pathway analysis.

Service Advantages

High Throughput

Leveraging HPLC and mass spectrometry, a large number of proteins can be identified and quantified in a short period of time.

High Efficiency

Bioinformatics tools can mine, integrate, and visualize large datasets.

Diversity

Suitable for various organisms and cell types, cellular proteomics analysis can be used to study protein changes under different environments, disease states, or genetic mutations.

Systematic

By comprehensively revealing the expression, modification, and interaction of all proteins within cells, it provides systematic and holistic information for biological research.

Sample Submission Requirements

In the technical report, MtoZ Biolabs will provide you a detailed technical information, including:

Experimental ProceduresRelevant Mass Spectrometry ParametersCellular Proteomics Analysis DetailsMass Spectrometry ImagesRaw Data

Cell surface proteins form a unique class of proteins, playing critical roles in managing cell function and facilitating communication between the cell and its environment by transporting metabolites, ions, and other solutes. MtoZ Biolabs provides integrate Cell Surface Proteomics Service.

Cell surface proteins vary across different cell types and can even change within a specific cell type under normal or diseased conditions. Consequently, cell surface proteins have numerous significant applications, such as distinguishing cell phenotypes and disease states and aiding research and development for prognosis and therapeutic targets. The cell surface contains key biomarkers and potential drug targets for various diseases. Approximately 70% of current biopharmaceuticals in development target cell surface proteins, underscoring the importance of studying surface proteins and their potential role in developing novel cancer therapies.

MtoZ Biolabs provides specialized and comprehensive cell surface proteomics services for identifying and quantifying the cell surface proteome, encompassing the entire mass spectrometry analysis workflow, from cell surface protein extraction to protein identification and quantification.

Leveraging biotin affinity purification technology, MtoZ Biolabs has optimized protocols for cell surface protein purification and enrichment, enabling high-quality extraction and enrichment of cell surface proteins for subsequent proteomic analysis. The company uses the Orbitrap Fusion Lumos mass spectrometer, the latest high-resolution, high-sensitivity instrument from Thermo, in combination with nanoLC chromatography for comprehensive cell surface proteomics analysis. With robust bioinformatics capabilities and extensive experience in bioinformatics analysis, our technicians can proficiently utilize various bioinformatics analysis tools to conduct in-depth data analysis and mining, allowing for effective screening of potential biomarkers.

Exosomes are one of the common extracellular vesicles (EVs) with diameters ranging from 30 to 150 nm. They primarily originate from the invagination of lysosomal particles into multivesicular bodies, which are released into the extracellular matrix, such as tissues and various biological fluids including serum/plasma, cerebrospinal fluid, and urine (in vivo), after the outer membrane of the multivesicular body fuses with the cell membrane. Additionally, exosomes can also exist in vitro under certain conditions in culture media. EVs play an important role in various physiological processes, and compared to general EVs, the function of exosomes in the field of clinical biomarkers is still unclear. With the development of exosome separation technologies, interest in this field is growing, and it is anticipated that more detailed studies on the function and applications of exosomes in disease pathologies will emerge. MtoZ Biolabs provides integrate Exosome Proteomics Service.

Exosomes primarily contain a complex mixture of proteins, lipids, RNA, etc. The content and function of exosomes from different biological fluids vary, and compared to other contents, proteins play a crucial role in biological functions. Therefore, proteomic analysis of exosomes from different biological samples is significant for the discovery of biomarkers and clinical diagnosis.

MtoZ Biolabs provides professional and comprehensive exosomal proteomics analysis services, capable of identifying and quantifying proteins in exosomes. By integrating bioinformatics techniques for the analysis of proteomic data, the detection of biomarkers can be achieved.

Figure 1. Saliva and Serum Exosome Proteomics Workflow (Y. Sun et al., 2017)

Service Advantages

One-stop Exosome Proteomics Analysis

We can complete the entire process including exosome separation to protein identification and quantification. It is possible to separate exosomes from various types of samples, including plasma/serum, cerebrospinal fluid, saliva, snot, urine, bile, and cell culture media.

Advanced Facilities and Optimized Operation Procedures

MtoZ Biolabs uses the currently highest resolution and sensitivity mass spectrometer, the newly introduced Obitrap Fusion Lumos by Thermo company, combined with nanoLC chromatography for exosomal proteomics analysis. MtoZ Biolabs has extensive experience in the field of exosome proteomics analysis and can provide customized services.

Biomarker Detection through Bioinformatics Analysis

Our professional bioinformatics analysts are proficient in using various bioinformatics analysis tools, providing in-depth analysis and data mining.

For mass spectrometry-based proteomic analysis, animal samples typically include fresh tissues, blood, urine, and cells. Paraffin-embedded (FFPE) samples, especially those used clinically, offer the advantages of well-documented medical histories, accurate diagnoses, and comprehensive clinical data. They can be stably stored at room temperature for extended periods, making them highly valuable for disease research. Therefore, proteomic research on FFPE samples is of great importance in understanding disease mechanisms, identifying biomarkers, and studying rare diseases. MtoZ Biolabs provides integrate Paraffin Embedded Sample Proteomics Service.

MtoZ Biolabs uses Thermo Fisher's Orbitrap Fusion Lumos mass spectrometry platform combined with nanoLC chromatography to provide comprehensive technical solutions for FFPE sample proteomic analysis. Simply communicate your research objectives and send us your samples, and we'll handle the rest, including protein extraction, protease digestion, peptide separation, mass spectrometry analysis, raw mass spectrometry data processing, and bioinformatics analysis.

Analysis Workflow

Deliverables

In the technical report, MtoZ Biolabs will provide you with a detailed technical information, including:

Experimental ProceduresRelevant Mass Spectrometry ParametersMass Spectrometry ImagesRaw DataBioinformatics Analysis

The distribution and compartmentalization of proteins encoded by intracellular nucleic acids enable specific cellular functions. Therefore, analyzing the protein expression of their subcellular compartments is a practical and necessary functional assay for cellular proteomics. MtoZ Biolabs provides integrate Subcellular Proteomics Service.

The powerful combination of subcellular fractionation and mass spectrometry protein identification technologies has helped us develop methods that can characterize subcellular-level proteomics. Subcellular proteomics, built on decades of biochemical research, has already enabled subcellular fractionation. Advances in techniques such as density, size, and charge separation also allow subcellular structure separation to limit the level of contaminants. Coupled with mass spectrometry, high-level analysis at the subcellular level containing 500-4000 proteins can be achieved. Compared to the whole-cell proteome, which contains 12, 000-40, 000 proteins, subcellular proteomics reduces the complexity of exploring the proteome.

Combining subcellular fractionation with proteomic analysis seems to be an effective approach to simplify the complex proteins extracted from cells or tissues and facilitate the detection of low-abundance proteins. However, realizing subcellular proteomic analysis involves each complex step in sample preparation, analysis, and verification, and only by ensuring the quality of each step can the potential of subcellular proteomic analysis be fully utilized.

MtoZ Biolabs uses Thermo Fisher's Orbitrap Fusion Lumos mass spectrometry platform combined with nanoLC chromatography to provide subcellular proteomic analysis services, including qualitative and quantitative analysis of membrane proteomes, mitochondrial proteomes, histones, chloroplast proteomes, and more.

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

Experimental ProceduresRelevant Mass Spectrometry ParametersMass Spectrometry ImagesRaw DataBioinformatics Analysis

Targeted proteomics is a scientific discipline focused on the quantitative study of specific target proteins, building on the foundation of general proteomics. Unlike non-targeted proteomics, this approach selectively chooses ions for the collection of mass spectrometry data, enabling the analysis of numerous samples with enhanced accuracy, sensitivity, and reproducibility. Targeted proteomics is applicable to a range of studies including post-translational modifications, protein conformations, protein-protein interactions, dynamics, and the systemic investigation of metabolic and signaling pathways. MtoZ Biolabs provides integrate Targeted Proteomics Service.

multiple reaction monitoring (MRM) and parallel reaction monitoring (PRM). MRM, also referred to as selected reaction monitoring (SRM), employs triple quadrupole mass spectrometry (QQQ-MS) to capture signals conforming to predefined ion criteria, facilitating both relative and absolute quantification of target proteins. PRM extends the capabilities of MRM by integrating the high selectivity of quadrupoles with the superior resolution and precision of the Orbitrap technology. It measures all fragment ions following the fragmentation of selected parent ions, offering improved detection sensitivity and reduced susceptibility to interference.Targetedproteomicsquantitativetechniquesmainly include

MtoZ Biolabs leverages a high-throughput mass spectrometry platform to offer targeted proteomics analysis services, which include:

Parallel Reaction Monitoring (PRM) ServicesMRM/PRM-Based Quantitative Proteomics AnalysisAbsolute Quantification Analysis AQUA

Gel electrophoresis is an important protein analysis method, which includes multiple techniques such as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing (IEF). SDS-PAGE is the most commonly used technique in protein analysis, providing a simple way to estimate the molecular weight of proteins, evaluate sample complexity or preparation purity, and monitor products obtained during chromatography or other purification processes. Additionally, ICH Q6B requires electrophoresis profiles for biopharmaceutical analysis. MtoZ Biolabs provides integrate Protein Gel and Imaging Analysis Service.

Services at MtoZ Biolabs

Gel electrophoresis has been used in protein research for many years, separating proteins based on differing physicochemical parameters. MtoZ Biolabs provides a variety of protein gel and image analysis services.

Protein Separation Based on SDS-PAGE

SDS-PAGE is often used in protein expression analysis. It can provide electrophoresis profiles of protein samples and analyze or purify samples in combination with other mass spectrometry-based protein identification services for proteomic analysis of complex biological samples.

1D SDS-PAGE and IEF Services

1D SDS-PAGE separates proteins based on differences in molecular weight. SDS-PAGE is widely used in protein analysis, including the determination of protein size, protein identification, and sample purity analysis.

2-DE Services

Two-dimensional electrophoresis (2-DE) is a method to separate complex protein mixtures. 2-DE first uses IEF, an electrophoresis technique for protein separation based on isoelectric points (pI), to separate proteins with similar sizes. SDS-PAGE is then used for further separation. MtoZ Biolabs provides 2-DE analysis services. High-resolution 2-DE can analyze protein species and subtype distribution, host cell protein analysis, and quantitative comparisons of cellular states.

2D BN/SDS-PAGE Complex Analysis Services

In biomembranes, many proteins are composed in complexes. MtoZ Biolabs can comprehensively analyze the subunits of these protein complexes using 2D Blue Native (BN)/SDS-PAGE. In 2D BN/SDS-PAGE analysis, samples are separated in the 1D by blue native polyacrylamide gel electrophoresis (BN-PAGE) and then in the 2D using SDS-PAGE.

WB and Electrotransfer Services

Western blot (WB) is another commonly used protein analysis method for qualitative and semi-quantitative protein analysis. In this method, proteins are first separated by 2-DE and then transferred onto a carrier membrane through electrotransfer. WB can be used for qualitative detection of individual proteins and protein modifications.

Service Advantages

Compatible with Multiple Techniques, Including Molecular Weight Measurement, Impurity Characterization, Protein Expression Research, etc.Suitable for Various Sample Types, Including Body Fluids, Tissues, Blood Samples, Cell Cultures, Microorganisms, Plants, etc.Multiple Gel Size Options, Including 87 cm, 2330 cm, 4030 cm, and 6030 cmExperienced Technicians can Provide Diverse, Customizable Protein Gel and Image Analysis Services

MtoZ Biolabs provides comprehensive protein gel analysis services, including SDS-PAGE, IEF, and Native PAGE.

1D SDS-PAGE Services

Polyacrylamide gel electrophoresis (PAGE) is a widely used technique for separating macromolecules, including DNA, RNA, and proteins. In this process, charged ions migrate under an electric field. The mobility of charged molecules is proportional to their net charge and the resistance of the solution they pass through. Sodium dodecyl sulfate (SDS), an anionic detergent, imparts a net negative charge to polypeptide chains over a broad pH range. Polypeptide chains bind to a consistent amount of SDS relative to their molecular weight, and the negative charge of the SDS disrupts most of the protein's complex structures, effectively denaturing them.

When a protein sample is added to the gel loading well, larger proteins may not have entered the gel yet, while smaller proteins may have already migrated beyond it. To achieve better separation, the gel is divided into two layers: a stacking gel and a separating gel. The separating gel, located at the bottom, is wider while the stacking gel above it is narrower. The electrophoresis buffer, made with glycine at pH 8.3, interfaces with the stacking gel at pH 6.8 and the separating gel at pH 8.8. During electrophoresis, negatively charged molecules migrate in the same direction. Because only about 10% of glycine ions carry a negative charge at pH 8.3, the current is mainly carried by Cl- ions in the stacking buffer, which migrate ahead of the proteins at the bottom of the stacking gel. Protein molecules coated with SDS are trapped between glycine ions above and Cl- ions below, forming a narrower band than initially loaded.

As proteins migrate into the separating gel, glycine in the separating gel buffer at pH 8.8 becomes negatively charged and migrates rapidly, closely aligning with Cl- ions, with proteins following just behind. At this stage, proteins migrate into the separating gel along the path of Cl- and glycine anions, carrying the current. Proteins become rod-like, negatively charged high-molecular-weight polymers with similar hydrodynamic properties, and their mobility depends only on molecular weight.

SDS-PAGE is widely used in proteomics, including protein molecular weight determination, protein identification, sample purity analysis, disulfide bond identification, and protein quantification.

IEF Analysis Services

Isoelectric focusing (IEF) is an electrophoresis technique used to separate proteins based on their isoelectric points (pI). When pI equals pH, the protein's net charge is zero, so the protein does not migrate in the electric field. In IEF, separation occurs in polyacrylamide or agarose gel containing a mixture of ampholytes that migrate in the electric field and form a pH gradient in the gel. Protein molecules are concentrated in a medium with a pH gradient, generating a positively charged anode and a negatively charged cathode. Charged protein molecules move toward the opposite electrode until they reach a point where the surrounding pH matches their pI, at which point their net charge becomes zero, and they stop migrating.

Proteins with the same pI concentrate in fixed pH bands. Each protein is located at the pH gradient corresponding to its pI. IEF has extremely high resolution, allowing proteins differing by a single charge to be separated into distinct bands.

Figure 2. IEF Analysis

In biological membranes, many proteins form complexes. MtoZ Biolabs provides comprehensive subunit analysis of these protein complexes using 2D blue native (BN)/SDS-PAGE. In 2D BN/SDS-PAGE analysis, samples undergo 1D separation through BN-PAGE, followed by 2D separation via SDS-PAGE. MtoZ Biolabs provides integrate 2D Blue Native/SDS-PAGE Protein Complex Analysis Service.

Protein complexes organize and maintain cellular and organelle functions at all levels of complexity in both time and space, including cell development and division, transcription and translation, respiration and photosynthesis, transport, and metabolism. Protein complexes may exhibit partial and transient changes and show varying stability, making their identification and analysis challenging. BN-PAGE is a high-resolution separation technique for protein complexes. It can be used to determine the size, relative abundance, and subunit composition of protein complexes.

In 2D BN-PAGE analysis, protein or protein complex samples are separated in 1D BN-PAGE under native conditions. Coomassie brilliant blue dye carries a negative charge and binds non-specifically to all proteins. Unlike detergents, coomassie brilliant blue preserves the structure of protein complexes. Its binding also reduces the tendency for protein aggregation during electrophoresis. The electrophoretic mobility of the samples depends on the negative charge of the bound coomassie brilliant blue as well as the size and shape of the complex. After separation by BN-PAGE, protein and protein complex samples are denatured in-gel with SDS and then separated using 2D SDS-PAGE.

The combination of BN-PAGE and SDS-PAGE enables monomeric proteins to migrate along a hyperbolic diagonal due to the gradient gel in 1D and linear gel in 2D. Components of protein complexes appear below this diagonal. Proteins representing the subunits of the same protein complex can be found in a vertical line in 2D, while multiple spots of the same protein along the horizontal line indicate that the protein is simultaneously present in several different protein complexes.

2D BN/SDS-PAGE analysis provides valuable information regarding the size, quantity, protein composition, stoichiometry, and relative abundance of samples, making it a powerful tool for studying protein complexes.

About Us

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

Two-dimensional gel electrophoresis (2-DE) is a powerful and widely used method. It combines IEF and SDS-PAGE. First, proteins are separated by their isoelectric point (pI) in IEF, and then by SDS-PAGE based on molecular weight. Each protein spot on the two-dimensional gel can be eluted and identified by high-throughput mass spectrometry. Therefore, 2-DE is particularly suitable for comparing protein profiles between different tissues, different conditions, or between control and experimental samples. MtoZ Biolabs provides integrate Two-Dimensional Gel Electroph

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

Glycosylation modification not only affects the spatial conformation, biological activity, transport, and localization of proteins, but also plays a crucial role in specific biological processes such as molecular recognition, cell communication, and signal transduction. MtoZ Biolabs provides integrate Glycoprotein Analysis Service.

With the rapid development of protein drugs, monoclonal antibody (mAb) drugs develope rapidly. The glycosylation of mAb drugs has varying degrees of influence on the stability, efficacy, and safety of mAb. Therefore, comprehensive characterization of glycosylation modification is of great significance. For mAb drugs, macromolecular protein drugs, it is essential to evaluate their primary or secondary structure that can affect their efficacy and safety. Hence, qualitative and quantitative studies of glycosylation modifications of mAb drugs are crucial for drug research and development, improvement, and safe use.

Figure 1. Glycoprotein Analysis Service by MtoZ Biolabs

Services at MtoZ Biolabs

Glycan Type AnalysisGlycosylation Site AnalysisGlycosylation Sites and Glycoform Analysis

About Us

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

For the relative quantification of glycan types in glycosylated proteins, MtoZ Biolabs provides glycan type relative quantification services based on MALDI-TOF/LC-MS. Due to the diversity of O-glycosylation modifications and the absence of common enzymes that can cleave all O-glycans, our glycan type analysis currently focuses only on N-glycosylated proteins. MtoZ Biolabs provides integrate Glycan Profile Analysis Service.

The strategy for glycan type analysis is as follows: all N-glycans in the target protein are digested by PNGase F enzyme, followed by labeling N-glycans with 2-AA, and specifically enriching N-glycans through an SPE column. Depending on the complex glycosylation of the target protein, MALDI-TOF-TOF or LC-MS/MS is used to detect 2-AA-labeled N-glycans, and GlycoWorkbench software (CFG database) is used to identify N-glycans and analyze the relative abundance of glycan types based on the mass spectrometry peak areas of the glycans.

Service Advantages

Relatively Short Identification and Analysis TimeHigh Sensitivity and Good Reproducibility

Applications

Identification of N-glycan Types in Glycosylated ProteinsAnalysis of the Relative Abundance of N-glycan Types in Glycosylated Proteins

Sample Submission Requirements

If you provide tissue samples, please transport them on dry ice.If you provide protein samples, please transport protein extracted from ordinary tissue and cell lysis buffers.Sample Transport: Please use an ample amount of

The analysis of glycosylation sites in glycosylated proteins is of great importance for studying protein structure and function.MtoZ Biolabs provides integrate Glycosylation Site Analysis Service.

MtoZ Biolabs provides a glycoform relative content analysis service based on LC-MS/MS. First, various enzymes are used to digest the protein, then in heavy-labeled H218O, PNGase F or various O-glycosidases are used to enzymatically release the glycans from the target protein. At this point, the glycosylated peptides will carry a 18O, and by searching the database, the glycosylation modification sites can be identified.

Service Advantages

Relatively Short Identification and Analysis TimeHigh Sensitivity and Good Reproducibility

Applications

Identification of N-Glycosylation Sites in Glycosylated ProteinsIdentification of O-Glycosylat

Protein glycosylation not only influences the protein's spatial configuration, biological activity, transportation, and localization, but also plays a crucial role in specific biological processes such as molecular recognition, cell communication, and signal transduction. Therefore, the analysis of glycosylation sites and their glycoforms is of great significance. MtoZ Biolabs provides integrate Glycosylation Sites and Glycoform Analysis Services.

MtoZ Biolabs offers a glycosylation site and glycoform analysis service based on LC-MS/MS. We typically start by selecting various enzymes to digest the target protein to cover as much of the protein sequence as possible. After cleaving the glycans by enzymes, the glycosylation sites in the protein are analyzed. Once the glycosylated peptides are identified, the target protein is redigested and the glycans on the peptides are retained. During LC-MS/MS analysis, we use a combination of ETD, HCD, and CID to obtain as much secondary fragmentation information of the peptides as possible. Through software analysis and manual identification of the secondary spectra of glycosylated peptide fragments, accurate information about the glycosylation sites and their glycoforms of the target protein is obtained.

Figure 1. Glycosylation Sites and Glycoform Analysis Process

Service Advantages

Accurate Analysis of the Glycoforms Corresponding to the Glycosylation Sites in the Target ProteinHigh Sensitivity and Good Reproducibility

Applications

Identification of the Glycoforms Corresponding to N-glycosylation Sites in Glycosylated ProteinsIdentification of the Glycoforms Corresponding to O-glycosylation Sites in Glycosylated Proteins

Sample Submission Requirements

If you provide tissue samples, please transport on dry ice.If you provide protein samples, please transport protein extracted from common tissues and cell lysis buffer.Sample Transport: Please use an ample amount of dry ice and opt for the fastest possible mailing method to minimize sample degradation during transportation.

Glycomics refers to the collection, analysis, and utilization of glycosidic biological data at the omics level. The profound biological effects of lipid and protein glycosylation make glycomics an important field in life sciences. N-glycome analysis (a comprehensive study of all glycans) is a complement to genomics and proteomics. Glycomics studies at the cellular or organismal level can provide a general overview of the overall glycosylation patterns of glycome, glycoprotein, lipid, or other type of biomolecules. Glycomics analysis can be used to compare differences between two glyco-engineered host strains in the production of therapeutic glycoproteins or to compare cancer data at different stages to discover new biomarkers. MtoZ Biolabs provides integrate Glycomic Profiling Service.

Services at MtoZ Biolabs

We can identify all N-glycans expressed by plasma/serum, cells, tissues, or organisms. All glycans linked to proteins will be digested and hydrolyzed by enzyme and separated by hydrophilic chromatography, followed by quantitative analysis by MALDI-TOF-MS.

Analysis Workflow

Glycoprotein AnalysisQuantitative GlycoproteomicsN-glycan Analysis ServiceO-glycan Analysis ServiceN-glycosylation Site AnalysisO-glycosylation Site AnalysisN-glycan Modification and Modification Site Analysis ServiceO-glycan Modification and Modification Site Analysis ServiceHILIC-UHPLC Analysis of N-glycan Bonds

About Us

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

Glycosylation modifications increase the complexity of protein molecular structure and the diversity of functions, not only affecting protein folding, structure, transportation, localization, and biological activity but also regulating various physiological processes such as cell growth, division, differentiation, recognition, and adhesion through specific recognition between sugars and proteins. Among these, N-glycosylated proteins are ubiquitously present on cell surfaces and various body fluids. Surface glycoproteins are considered key drug targets, for instance, glycoproteins in body fluids are major sources of biomarkers related to diseases. Proteomics of N-glycosylated proteins under various physiological and pathological conditions can elucidate the function of protein glycosylation. It is important for identifying new biomarkers in cancer prevention and diagnosis. MtoZ Biolabs provides integrate N-Glycan Modification and Site Analysis Service.

According to the substitutes of mannose residues, N-glycans can be roughly divided into three types: complex, high-mannose, and hybrid structures. The complex type includes fucose, sialic acid, and -linked galactose. A branch of the high-mannose core structure contains multiple mannose residues. In the hybrid type, mannose is in one branch of the core structure, and another branch contains a lactosamine sequence. General N-glycan site analysis strategy: isolating and enriching glycoproteins/glycopeptides, then using enzyme or chemical methods to separate the sugar chains from the glycosylated peptides, and performing mass spectrometry (MS) analysis of the released sugar chains or deglycosylated peptides to achieve analysis of the overall expression and changes of sugar chains and/or glycoproteins in the test sample.

Figure 1. Types of N-glycans

Services at MtoZ Biolabs

MtoZ Biolabs offers N-glycan modification and site analysis services, capable of identifying modification sites, specific glycan types, analyzing N-glycan modification types and locations, and resolving the structure and location of N-glycans, and can perform quantitative analysis of glycan modifications.

N-glycan chain analysis typically follows these steps:

Release of Attached Glycans by EnzymeDerivatization of Released Glycans Through Reduction Amination by Aromatic Amine and Alkyl Amine or PermethylationAnalysis of Glycan

Service Advantages

Good Specificity and High EfficiencyHigh-throughput Identification Capable of Detecting up to 10, 000 N-glycosylation SitesHigh Sensitivity and Good ReproducibilityRelatively Short Project Time

Sample Submission Requirements

If you are providing tissue samples, please transport them on dry ice.If you are providing protein samples, please transport protein extracted from normal tissues or cell lysis buffer.

Ensure sufficient dry ice for transportation and choose the fastest mailing method to minimize sample degradation during transport. If you have any questions about sample transportation, please contact us at any time.

MtoZ Biolabs has much analysis service experience in glycomics and can customize glycomics analysis service according to your specific needs.

Free project evaluation!

Deliverables

Experimental ProceduresMass Spectrometric ParametersMass Spectrometric ImagesRaw DataResults of N-glycan Modification and Site Analysis

About Us

MtoZ Biolabs is an integrated Contract Research Organization (CRO) dedicated to providing cutting-edge chromatography and mass spectrometry services for researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our team of experts and advanced technological platforms are designed to support your research projects with precision and reliability.

Glycosylation primarily refers to the enzymatic process of linking polysaccharides to proteins, lipids, or other organic molecules, and is the most common type of post-translational modifications (PTMs). Due to the complexity and isoform nature of the linked polysaccharides, glycan analysis presents significant challenges. As protein therapeutics such as monoclonal antibodies, glycoproteins, hormones, cytokines, and coagulation factors are increasingly used in clinical practice, the detection of protein N-glycosylation (also known as N-glycan, N-sugar) has received growing attention. MtoZ Biolabs provides integrate N-Glycan Profiling Service.

N-glycan is an oligosaccharide structure containing Man3-GlcNAc2, which is linked to the polypeptide chain of proteins through a glycosidic bond between the terminal N-acetylglucosamine (GlcNAc) and the NH2 group of asparagine. The sample preparation steps for most glycan analysis include glycoprotein digestion, deglycosylation, purification, labeling/derivatization, and SPE. Depending on the characteristics of the sample, other steps and techniques may also be used for sample preparation.

Utilization of MALDI-TOF-MS and/or UHPLC Techniques for Glycan and Type Analysis

Based on MALDI-TOF-MS Technique for N-glycan Analysis

MALDI-TOF offers high sensitivity in structural characterization of glycosylated compounds and allows for high-throughput detection. In the analysis of mixture N-glycans, MALDI performs the analysis by forming single charged ions, rather than using electrospray ionization. After releasing the N-glycans from the sample using N-glycosidase A or F, the sample is methylated for detection.

Based on HILIC-UHPLC-MS Technique for N-glycan Profiling Analysis

LC-MS-based N-glycan analysis is the current mainstream analytical method. Hydrophilic interaction chromatography (HILIC) is suitable for N-glycan analysis. N-glycans released by the PNGase F enzymatic reaction are derivatized using 2-aminobenzamide (2-AB), and the derivatized glycans are measured for large sample N-glycan analysis. Prior to fluorescence labeling, if there are detergents, non-volatile salts, or substances with free amino groups that might interfere with the derivatization reaction, the N-glycans can be purified using a TSK Amide-80 chromatography column. The labeled N-glycans are then analyzed by HILIC-UHPLC-MS.

MtoZ Biolabs has much analytical service experience in glycomics and can customize glycomics analysis services according to your specific needs.Free consultation is available!

Deliverables

Experimental ProceduresMass Spectrometric ParametersMass Spectrometric ImagesRaw DataN-glycan Analysis Results

Unlike N-glycans, O-glycans are smaller, highly diversified branched carbohydrate molecules that can attach to various protein structures. This characteristic leads to the complexity of the analysis, making it difficult to analyze O-glycosylation sites. The variability of glycosylation sites is a key indicator of cellular activity, as evidenced by the correlation between the reduction in serum protein glycosylation sites and the severity of congenital disorders of glycosylation (CDGs). Therefore, accurately determining site occupancy is crucial for understanding the impact of protein glycosylation on human health. MtoZ Biolabs provides integrate O-Glycosylation Site Analysis Service Biological Products.

The general steps in O-glycosylation site analysis include the proteolytic digestion of glycoproteins, enrichment and separation of glycopeptides via hydrophilic interaction liquid chromatography (HILC). Enriched O-glycopeptides can be analyzed using reversed-phase liquid chromatography-ion trap mass spectrometry (RPLC-ITMS).

MtoZ Biolabs has much analytical service experience in glycomics and can customize glycomics analysis services according to your specific needs. Free project evaluation!

Deliverables

Experimental ProceduresMass Spectrometry ParametersMass Spectrometry ImagesRaw DataDetailed Information on Identified Glycosylation Sites

The proteome comprises gene products expressed at varying levels. Preparing protein samples is a critical step in proteomics research because the quality and reproducibility of protein extraction significantly affect downstream mass spectrometry identification efficiency and accuracy. Identifying and quantitatively comparing low-abundance proteins presents a significant challenge in proteomics. MtoZ Biolabs provides integrate Sample Preparation Service.

The proteome is a highly complex system. Unlike the chemically homogeneous nature of DNA and RNA, proteins exhibit considerable heterogeneity, varying greatly in size, charge, hydrophobicity, and structure. High-abundance proteins, such as actin, tubulin, or human serum albumin, along with complement system proteins and antibodies, form essential components of the proteome. With appropriate pretreatment, the low-abundance regulatory proteins most relevant to cellular responses can be more effectively detected. Moreover, many proteins undergo post-translational modifications and contain various non-peptide groups, making protein separation highly challenging. Currently, there is no universal method for protein sample preparation in proteomics research. Preparation protocols differ based on sample origin, the physical properties and subcellular localization of the target proteins, protein abundance, and matrix effects.

MtoZ Biolabs provides sample preparation services and continuously develops customized methods to meet diverse client requirements.

Figure 1. Protein Sample Preparation Workflow

Services at Mtoz Biolabs

Tissue and Cell HomogenizationSample Purification (Removal of DNA, RNA, and Reduction of Disulfide Bonds)Removal of High-abundance ProteinsProtein ExtractionProtein Purification and EnrichmentProtein Digestion and Hydrolysis

Service Advantages

State-of-the-art Ultrafiltration Technology PlatformExperienced Technicians Skilled in Processing Various Sample TypesComprehensive Quality Control WorkflowProject Design Tailored to Target Protein Characteristics

Proteins found in nature vary in size from 5 kDa to 400 kDa. The protein hydrolysis process cuts proteins into shorter segments, namely peptides. Protein hydrolysis is a crucial step before peptide mass spectrometry identification analysis, providing a foundation for successful protein identification and characterization, as well as biomarker discovery. Although mass spectrometry (MS) can study intact proteins, smaller peptides are easier to identify proteins and improve protein coverage because protein coverage may decrease due to solubility and heterogeneity. Therefore, the most common proteomics methods often utilize site-specific cleavage sites to produce smaller peptide fragments. Smaller peptides are easier to separate and characterize using high performance liquid chromatography (HPLC) and HPLC-MS. MtoZ Biolabs provides integrate Protein Hydrolysis Service.

Services at MtoZ Biolabs

MtoZ Biolabs offers two methods for protein hydrolysis: (1) in-gel digestion (protein hydrolysis in polyacrylamide gel matrix); (2) in-solution digestion (protein precipitation in chloroform/methanol, then redissolved in urea). Proteins separated by 1D or 2D gel electrophoresis can be hydrolyzed directly in the gel. In-gel digestion is effective and reproducible but more labor-intensive and time-consuming. Compared to in-gel digestion, the in-solution digestion method requires less effort and less time, but proteins hydrolyzed in solution may have low resolubility, possibly leading to some sample loss.

Analysis Workflow

MtoZ Biolabs can provide both in-gel digestion and in-solution digestion services. The workflow of protein hydrolysis includes:

Prepare Lysis SolutionEnzymatic Digestion in Solution or GelDenature Proteins using Denaturants (e.g., Urea and Guanidine)Reduce Disulfide Bonds using DTTAlkylate Cysteine Residues with Iodoacetic Acid or IodoacetamideRemove Reagents and Exchange BufferDenature Overnight (18 Hours) with Trypsin or other Proteases in Ammonium Bicarbonate Buffer at an Appropriate PH and TemperatureStop Digestion by Adding Acidic SubstancesEnrich/Elute Proteins

The most demanding and crucial part of 2D gel electrophoresis technology is the analysis of the resulting protein images. The image data are compared and analyzed automatically. Comparative analysis of 2D gel images can reveal qualitative and/or quantitative changes in protein expression between individual samples or different groups. 2D gel imaging analysis provides various types of information to identify novel, missing, or modified proteins, quantify protein spots, determine the pI and Mr values of protein spots, perform enzymatic digestion, and conduct mass spectrometry analysis. MtoZ Biolabs provides integrate Two-Dimensional Gel Electrophoresis Images Analysis Service.

To analyze and compare complex 2D images, a scanner or camera is required to convert gel image data into digital information for in-depth analysis using computer software. During the image analysis process, scanning is generally performed using a pixel size of 100 mm. Although higher pixel resolutions can improve image quality, they result in files too large to process within a reasonable time frame and consume significant disk space.

In biopharmaceutical processes, particularly for protein and antibody drugs, charge heterogeneity analysis is crucial for ensuring quality control of bioproducts. Interactions between molecules with different charge can significantly impact the bioactivity, immunogenicity, half-life, and charge stability of drugs, ultimately affecting their efficacy, stability, and safety. Ion chromatography (IC) is a widely recognized technique for analyzing charge heterogeneity in biomolecules by leveraging the charge adsorption interaction between charged molecules and ion exchange resins. IC can effectively separate molecules with varying charge states, enabling accurate identification and measurement of charge characteristics in samples. This technology is a key tool for developing and ensuring the quality of vaccines, antibody drugs, enzyme preparations, and other protein-based medications. MtoZ Biolabs provides integrate Ion Chromatography-Based Charge Heterogeneity Analysis Service.

MtoZ Biolabs has developed an efficient charge heterogeneity analysis platform based on advanced ion exchange chromatography and professional data analysis methods. This platform provides comprehensive services, including routine charge heterogeneity analysis and in-depth charge profiling. Specific services include qualitative and quantitative analysis of charge variants, component analysis of charge heterogeneity, and assessment of how charge state affects drug stability and activity. We offer support across drug development, production, and quality control.Welcome to contact to learn more about our service!

Deliverables

Experimental ProceduresRelevant Mass Spectrometry ParametersDetailed Charge Heterogeneity Analysis by Ion ChromatographyMass Spectrometry ImagesRaw Data

Charge heterogeneity is a key attribute of biomolecules such as antibody, enzyme, and protein, affecting the molecule's stability, solubility, and biological activity. Changes of deamidation, oxidation, glycosylation, glycation, the N- or C-terminal, loss of three-dimensional structure, or reduction of disulfide bonds may lead to changes in charge heterogeneity. Since charge heterogeneity directly or indirectly affects the efficacy and safety of biomolecules, the analysis of charge heterogeneity occupies an important position in the research and development and production of biopharmaceuticals. MtoZ Biolabs provides integrate cIEF Based Charge Heterogeneity Analysis Service.

Capillary isoelectric focusing (cIEF) is an advanced technique specifically used for analyzing charge heterogeneity. It employs a capillary as the medium, creating a pH gradient and applying an electric field to achieve a refined analysis of charge heterogeneity by migrating protein or peptide molecules to their isoelectric points. Thereby . cIEF is widely used in the analysis of protein isoelectric points, charge heterogeneity, and protein modifications due to its high-efficiency and high-resolution capability.

MtoZ Biolabs has established a platform for the separation and comprehensive characterization analysis of charge variants, providing you with comprehensive and accurate cIEF charge heterogeneity analysis services. Whether you are focused on minute charge variants or need a comprehensive analysis of the entire molecule group, BTP can provide solutions that meet your needs. F.ree consultation is available

Deliverables

Experimental ProceduresRelevant Experimental ParametersDetailed Information on Charge Heterogeneity Analysis Based on cIEFElectrophoresis ImagesRaw Data

Collagen is a basic component of the extracellular matrix and the most abundant protein in the animal body, accounting for about 30% of the total protein. It is a glycoprotein, composed of three polypeptide -chains with Gly-XY repeating residues (Gly stands for glycine, X for proline, Y for hydroxyproline), and is renowned for its triple helix structure. Over 28 types of collagen have been identified, with Type I collagen being the most abundant. Collagen is commonly found in ligaments, tendons, skin, and bone tissues. Its mature, insoluble form provides extraordinary strength, making it essential for the mobility of organisms. Collagen also has biochemical functions that affect cell growth, proliferation, and differentiation. MtoZ Biolabs provides integrate Collagen Analysis Service.

Collagen can be degraded, converted, and absorbed back into the body, and has become one of the best-selling supplements. It has a wide range of uses in the medical and cosmetic fields.

Applications

Dermal filler: Collagen injections can fill shallow depressions on the skin, such as fine lines and wrinkles.Wound dressing: Collagen helps heal wounds by attracting new skin cells to the wound site.Periodontology: Collagen acts as a barrier, preventing fast-growing gingival tissue from developing into dental wounds, to help dental cells to regenerate.Vascular prosthesis: Donor collagen tissue grafts have been used to reconstruct arteries, assist in peripheral nerve regeneration, and produce vascular prostheses.

As a highly versatile material, collagen has been developed for wound care, burn care, orthopedic graft products, tissue engineering, hemostatic sponges, soft tissue augmentation injections, drug delivery carrier, ingredient in skin and hair care products. Depending on the raw material and the processing methods used for collagen, its chemical, physical, biological, immunological, or toxicological properties can change accordingly. Therefore, in the development or manufacturing process of biomaterials containing collagen, it is necessary to analyze collagen using the most appropriate testing methods to determine the characteristics of specific collagen materials and the residues during the production process.

Figure 1. Collagen Characterization

MtoZ Biolabs offers high-quality collagen analysis services based on multiple platforms, including physical and biochemical analysis and impurity analysis. Our company adheres to regulations and guidelines of NMPA, ICH, FDA, and EMA. A comprehensive quality system has been established, with data hot/cold/offsite backup, regular equipment calibration/periodic verification, software audit tracking, to provide you with one-stop collagen analysis services.

Services at MtoZ Biolabs

Collagen Characterization

(1) Collagen molecular weight

(2) Collagen content/purity (hydroxyproline assay)

(3) Terminal amino acid sequence

(4) Mass spectrometric peptide map

(5) Peptide coverage

(6) Higher-order structure

(7) Amino acid heterogeneity analysis

(8) Total protein content

(9) Moisture content

Biochemical Characterization of Collagen

(1) Solubility

(2) Isoelectric point

(3) Osmolarity

(4) SDS-PAGE analysis

(5) Total DNA

(6) Total lipid content

(7) Trypsin resistance

(8) Collagenase resistance

(9) GAGs abundance

(10) Elastin assay

(11) pH levels

Physical Characterization of Collagen

(1) Viscosity (mainly soluble samples)

(2) TEM or SEM-electron micrographs

(3) Mechanical testing (e.g., tensile strength)

(4) Optical microscopy

(5) Surface characterization

Collagen Impurity Identification

Hyaluronic acid (HA), also known as sodium hyaluronate, is a high molecular weight linear polymer with repeating disaccharide units. Each disaccharide unit comprises a D-glucuronic acid and a N-acetyl-D-glucosamine residue linked by glycosidic bonds. HA is widely distributed in the human body, particularly in the eyes, synovial fluid, and skin. It is known for its unique molecular structure and physicochemical properties, displaying a variety of important physiological functions such as lubricating joint, regulating vascular permeability, modulating proteins, and promoting wound healing. Importantly, HA is recognized as the best natural moisturizing substance found in nature, termed as the ideal natural moisturizing factor (NMF). It enhances skin metabolism, making the skin soft, smooth, elastic, and anti-aging, and acts as a good transdermal absorption promoter while moisturizing. MtoZ Biolabs provides integrate Hyaluronic Acid Quality Control Testing Service.

Currently, HA is extensively used in the cosmetics industry, biomedical industry, etc., making accurate testing of HA-containing new products crucial for developing new injectable soft tissue fillers, wrinkle fill gels, cosmetics, and medical devices such as tissue scaffolds. HA is also used in ocular solutions, enteral medications, nasal sprays, aerosols, and is a key active ingredient in products aimed at preventing skin aging (such as viscous supplements). Understanding the quantity, purity, physicochemical properties, chemical composition, and microbiological characteristics of HA is essential for supporting its research and development, monitoring raw material quality, and the manufacturing process.

MtoZ Biolabs have developed and validated a unique and accurate analytical service based on multiple platforms like ICP-MS, ICP-AES, GC-MS, IC, etc., to control the quality of raw materials and finished products containing HA. Additionally, we provide method development and validation, stability studies under controlled temperature and humidity, and leachables and extractables studies. MtoZ Biolabs focuses on product quality and safety, helping clients meet and exceed the quality, safety, and regulatory standards for HA products. MtoZ Biolabs HA quality control testing includes the analysis of the following characteristics.

Services at MtoZ Biolabs

Appearance, Form and ColorHA Dosage/ConcentrationBDDE (1, 4-butanediol Diglycidyl Ether), Lidocaine Hydrochloride Content DeterminationStructure Identification/ConfirmationStability DeterminationOrganic Volatile Impurities and Particulates, Water Content, Protein Content, Residual Chemicals or Solvents, Degradation Products DeterminationPhysicochemical Properties, such as pH, Osmolarity, Viscosity, and Particle SizeHeavy Metals AnalysisCompatibility Testing

MtoZ Biolabs is committed to providing professional service like quality control testing and project validation for the bio/pharmaceutical and medical device industries. The laboratory complies with regulations and guidelines from NMPA, ICH, FDA, and EMA. With a complete quality system, cold/hot remote data backup, regular equipment calibration/interim checks, software audit tracking, we provide one-stop solutions and technical services to support bioproduct research and development, application registration, and production release. Our HA quality control testing platform, developed and validated, offers a range of chemical, biochemical, microbiological, and physicochemical testing services to assist you in quality control, ensuring the.

CUT&Tag (Cleavage Under Targets and Tagmentation) is an innovative tool dedicated to DNA-protein interaction research, primarily used to identify transcription factor or histone modification binding sites on a genome-wide scale. The CUT&Tag analysis service utilizes cleavage and transposase tagging technology to precisely map the binding sites of transcription factors or histone modifications within the genome, facilitating research into DNA-protein interaction mechanisms. It provides an accurate solution for genomic and epigenetic studies. Its high sensitivity, low background noise, and exceptional adaptability to low-input samples make it stand out in the field of epigenetics research.

Principles of CUT&Tag Analysis

In genomic and epigenetic studies, exploring the mechanisms of DNA-protein interactions is key to understanding gene regulation. Transcription factors regulate the recruitment of RNA polymerase and the initiation of gene transcription by binding to specific DNA sequences in promoter or enhancer regions. The efficiency of their binding determines the spatiotemporal pattern of gene expression. Histones, as the core components of chromatin, regulate chromatin accessibility and DNA availability through post-translational modifications (e.g., acetylation and methylation), thereby influencing the binding of transcription factors to DNA. Such interactions can either activate gene expression or induce silencing. Protein-DNA interaction analysis elucidates the synergy between transcription factors and histone modifications in gene regulation, providing crucial insights into epigenetic mechanisms and disease pathogenesis.

Schematic Diagram of the Protein-DNA Interactions Observed in the Crystal Structure

CUT&Tag employs Protein A/G-Tn5 fusion proteins to directly bind target proteins, eliminating the need for traditional crosslinking steps, thereby significantly reducing background noise and achieving high-resolution detection of binding sites. Compared to traditional ChIP-seq, CUT&Tag is more suitable for low-input samples, particularly excelling in studies of histone modifications and transcription factor binding sites. It serves as a powerful tool for uncovering gene regulatory mechanisms, offering efficient solutions for disease research and drug development.

Service at MtoZ Biolabs

MtoZ Biolabs provides the CUT&Tag analysis service leveraging an advanced Protein A/G-Tn5 fusion protein system and high-throughput sequencing platforms. We specialize in analyzing protein-DNA interactions, with a focus on transcription factor binding sites and the genome-wide distribution of histone modifications. Our CUT&Tag analysis service encompasses the entire workflow including sample preparation, sequencing, and data analysis while offering comprehensive genome annotation, functional analysis, and pathway mapping. We empower clients to unravel gene regulatory networks with precision, advancing epigenetics, disease research, and drug discovery.

1. Sample Preparation and Bead Binding

ConA beads are used to bind glycoproteins on the cell membrane, allowing for the extraction of nuclei or the direct use of nuclear samples. Simultaneously, digitonin is applied to permeabilize the cell membrane, enhancing nuclear membrane permeability and facilitating subsequent antibody binding.

2. Primary and Secondary Antibody Binding

Specific antibodies (primary antibodies) targeting the protein of interest are added and incubated (e.g., -H3K27me3 as a positive control and isotype control IgG as a negative control), followed.