Antibody Glycosylation Detection

Antibody glycosylation detection is a technology used to analyze the glycan structure and composition on antibody molecules. Glycosylation is a common post-translational modification process of proteins, which is crucial to the function of antibodies, including their stability, recognition, and antigen binding ability. In the biopharmaceutical industry, especially in the process of developing and producing monoclonal antibody drugs, the detection of antibody glycosylation is an essential step in ensuring product quality and efficacy.

 

Antibody glycosylation usually occurs in the Fc region of the antibody. The type and structure of such glycosylation can affect the function of the antibody, such as regulating immune responses, enhancing or reducing cytotoxicity, and changing half-life, etc. Different forms of glycosylation (such as high mannose type, complex type, or hydrolyzed type glycans) can lead to differences in antibody function.

 

Methods for Antibody Glycosylation Detection

1. Mass Spectrometry (MS)

Analyzing the glycosylation site and glycan composition of proteins or peptides by measuring the spectrum. This is a very sensitive and accurate method that can provide detailed glycan structural information.

 

2. Capillary Electrophoresis (CE)

Separating glycosylated forms based on differences in electrophoretic migration rates. This method is fast and has a high resolution.

 

3. Enzyme-Linked Immunosorbent Assay (ELISA)

Detecting and quantifying glycosylated structures using specific anti-glycan antibodies.

 

4. Fluorescent Labeling and Flow Cytometry

Labeling glycans with fluorescence, then detecting and analyzing them using flow cytometry.

 

When performing glycosylation analysis, it is usually necessary to appropriately process the antibody samples, such as protein purification, enzyme digestion, glycan release and labeling for detection. In the process of biopharmaceutical production, glycosylation analysis is an indispensable quality control step, ensuring the consistency, stability, and biological activity of the drugs.