Common Methods for Glycan Analysis of Monoclonal Antibodies
Monoclonal antibodies (mAbs) are produced by a single clone of B lymphocytes and bind with high specificity to a particular antigen. Beyond their protein structure, the glycan chains of monoclonal antibodies significantly impact their function, biodistribution, immunogenicity, and stability. Therefore, glycosylation analysis of monoclonal antibodies is a critical step in drug development.
The following are common methods for glycosylation analysis of monoclonal antibodies:
1. Capillary Electrophoresis-Mass Spectrometry (CE-MS):
This method is particularly suitable for analyzing N-glycans. Capillary electrophoresis is used for glycan separation, while mass spectrometry is used for structural identification.
2. Liquid Chromatography-Mass Spectrometry (LC-MS):
This can be used to analyze both N-glycans and O-glycans. Glycans are first released from the antibody using enzymatic or chemical methods, then separated by liquid chromatography and structurally identified by mass spectrometry.
3. Fluorescent Labeling and HPLC Analysis:
Glycan chains are first released and reacted with a fluorescent label, followed by separation and quantification using HPLC.
4. Lectin Blotting:
Specific lectins bind to specific glycan structures and can be used to identify particular glycan structures.
5. NMR (Nuclear Magnetic Resonance):
Provides detailed information on glycan structures, but requires large sample amounts and has high technical demands.
6. Enzyme-Linked Immunosorbent Assay (ELISA):
Uses specific glycan-binding proteins or antibodies to detect and quantify glycan chains.
Each technique has its advantages and limitations. The choice of method often depends on the purpose of the analysis, the available sample amount, the required sensitivity and resolution, and the available instrumentation. In different stages of drug development, multiple techniques may be needed to comprehensively evaluate the glycosylation heterogeneity and consistency of the product.
Figure 1
BiotechPack, A Biopharmaceutical Characterization and Multi-Omics Mass Spectrometry (MS) Services Provider
Related Services:
Glycosylation Site and Glycan Structure Analysis
N-Glycan Modification and Modification Site Analysis Services
How to order?
