Comprehensive Glycosylation Analysis Service

Glycosylation is a very important type of protein post-translational modification (PTM). It is estimated that over half of eukaryotic proteins undergo glycosylation, with glycoproteins playing pivotal roles in many biological processes and have important cellular functions.The two main types of protein glycosylation are N-glycosylation (glycans bound to asparagine) and O-glycosylation (glycans bound to serine or threonine). Glycosylation is the process by which proteins or lipids are attached to sugars under the control of enzymes, starting at the endoplasmic reticulum and ending in the Golgi apparatus. Glycosyltransferases transfer sugars to proteins, and form glycosidic bonds with amino acid residues on proteins. Proteins undergo glycosylation to form glycoproteins.Glycosylation has an important role in modifying proteins, regulating proteins, and helping protein folding functions. It has an important impact on the properties and functions of proteins, and is a new technology that can change the properties of proteins, which is of great biological significance.

 

Mass spectrometry can be used to detect and analyze glycosylation sites and glycoforms. MtoZ Biolabs can provide LC-MS/MS-based protein glycosylation analysis, including glycosylation sites and corresponding glycoform analysis services.We usually first select a specific enzyme for enzymatic digestion, and then desalt the enzymatically digested peptides for glycosylated peptide enrichment, which can provide high enrichment efficiency and more comprehensive characterization of glycoproteins. For LC-MS/MS analysis, HCD/SCE or ETD and HCD combined fragmentation is used to obtain as much secondary fragmentation information as possible. Through software analysis and manual proofreading of the secondary spectrum of glycosylated peptides, accurate information about the glycosylation site and the glycoform on protein was obtained.

 



 

Analysis Workflow

1. Protein extraction.

2. Protease digestion to obtain peptides.

3. Peptide desalination and glycopeptide enrichment.

4. LC-MS/MS professional glycosylation analysis.

5. Use professional software for N-glycosylation analysis and O-glycosylation analysis.

 

Glycosylation schematic: N glycan-protein linkage (left), O glycan-protein linkage (right).

 



 

Service Advantages

1. There is no limit to the type of sample.

2. Select the appropriate sample pretreatment according to the sample type.

3. Provide the latest database for analysis.

4. Provide (N-Glycan, O-Glycan) glycoforms, glycosylation sites and correspondence between glycoforms and sites.

5. Quantitative glycosylation: On the basis of identification, quantitative methods are used to measure the abundance of glycosylation modifications in proteins.

6. Functional research: Functional research on key glycosylation modification proteins to reveal the regulatory role of glycosylation modification in cell signaling and biological processes.

 

Sample Results

 



Figure 1. Five Types Glycoforms with a High Proportion of Glycoforms

 

Sample Submission Requirements

1. Sample volume: the total amount of protein should be greater than 500 μg.

2. Sample status: solution or dry powder is acceptable.

3. Sample type: purified protein or protein complex.

 

Services at MtoZ Biolabs

 



 

Applications

Biologics, including protein drugs, antibody drugs, vaccines, collagen, purified proteins and protein complexes.

 

FAQ

Q1: In addition to the N-X-S/T(X!=P) motif, is there any other motif type for N glycosylation?

Yes, non-standard motifs such as N-X-C.

 

Q2: Does O glycosylation have the same core law of pentaccharide as N glycosylation?

There is currently no specific law for O glycosylation.

• • Oligosaccharide Chain Structure Analysis Service

  Glycosylation has significant impacts on the efficacy, stability, and immunogenicity of biopharmaceuticals. Among these, the oligosaccharide spectrum and glycan structure are important indicators for assessing the characteristics of biopharmaceuticals. Oligosaccharides, as important carbohydrates, participate in various cellular life processes, such as protein folding and signal transduction. In addition, oligosaccharides often bind with proteins to form glycoproteins.

• • Glycosylated Peptides Analysis Service

  Glycosylation is a common post-translational modification (PTM) of proteins, which significantly affects the function and stability of proteins. Glycosylated peptides refer to forms where glycans are attached to peptide segments, containing information on the amino acids of the peptides, complete glycan information, and the binding sites of glycans and peptides.

• • Glycan Profiling Service

  Glycosylation is an important post-translational modification of proteins. Based on the different linkage modes between the glycan and peptide chains, protein glycosylation can be divided into N-glycosylation and O-glycosylation. N-glycosylation involves the linkage between the reducing end of N-acetylglucosamine (GlcNAc) and the nitrogen atoms on certain asparagine (Asn) side chain amides in the peptide chain.

• • N-Glycan Analysis Service

  As the pharmaceutical industry evolves, biopharmaceutical characterization has gradually become a hot research within the pharmaceutical sector. Glycosylation significantly influences the efficacy, stability, and immunogenicity of protein dru, and the investigation of glycan structures plays a pivotal role in biopharmaceutical characterization processes.

• • Glycosylation Site Detection Service

  MtoZ Biolabs uses both MALDI TOF MS and nano LC-ESI-MS/MS technology to provide efficient and accurate glycosylation analysis of multiple biopharmaceuticals