Glycosylation site analysis
Overview
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.
Glycosylation Site Analysis of Antibody
Glycosylation Sites Analytical Workflow
(1) Digestion of biopharmaceutical samples by trypsin or other appropriate proteases
(2) Separation of N-glycopeptides and O-glycopeptides through chromatographic column
(3) Deglycosylation treatment
(4) Peptide analysis by MALDI-TOF MS, or ESI-MS/MS
(5) Analysis of the N/O-glycosylation site based on the mass spectra results
(6) Comparison of the analytical results of glycosylation sites of different batch samples
Reports
• Experiment procedures
• Parameters of liquid chromatography and mass spectrometer
• MS raw data files
• Glycosylation site results
• Bioinformatics analysis
Related Services
Molecular Weight Analysis
N/C Terminal Sequencing
Peptide Mapping
Amino acids composition
Glycan Profiling
Disulfide bond analysis
Antibody C-terminal variation analysis
Purity analysis SDS-PAGE
Purity analysis SEC and RPLC
Host Cell Protein (HCP) Analysis
Antibody-Drug Conjugate (ADC) Analysis
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