Anti-Glycan Antibodies in Characterizing Cell-Surface Glycoconjugates

The glycoconjugates are essential components of the glycocalyx of many cell types. They play a role in many functions, including osmoregulation, cell-cell recognition, binding of hormones, protection of cells from phagocytosis and dehydration, differentiation, defense, and ion transport. Characterizing the nature, spatial orientation, and distribution of cell-surface glycoconjugates is often performed using specific anti-glycan antibodies. Here, Creative Biolabs gives an introduction to the approaches and advantages of using anti-glycan antibodies for cell-surface glycoconjugate characterization.

Fig.1 Interaction of carbohydrate binding proteins and macromolecules with cell surface glycans presented on glycoproteins and glycolipids. (Oyelaran, 2009)

Approaches for Using Anti-Glycan Antibodies for Characterizing Cell-Surface Glycoconjugates

• Immunohistochemistry - Firstly, tissues are prepared and fixed for histological staining. Then, the fixed tissues are incubated with appropriate antibodies. The bound antibodies can be visualized by the use of secondary reagents, i.e. labeled secondary antibody. This approach has the advantage of revealing the spatial orientation of different glycans, their relative abundance, and whether they are intracellular and/or extracellular. This information is difficult to obtain by using any other approach.
• Flow cytometry and cell sorting - Firstly, cells are incubated with specific anti-glycan antibodies that are biotinylated and conjugated to fluorescently labeled streptavidin or directly fluorescently labeled. Then, cells bound with the antibodies can be identified with their fluorescence in the flow cytometer. The number of binding sites is correlated with the degree of fluorescence. Moreover, the specific membrane location of different glycans can be identified using confocal microscopy and electron microscopy.

Advantages of Using Anti-Glycan Antibodies for Characterizing Cell-Surface Glycoconjugates

• Useful for characterizing cell-surface glycans when a limited number of cells is available
• Useful for identifying unique glycan determinants and changes in their expression during cell differentiation
• Useful for identifying minor differences in protein glycosylation between different samples
• Useful for identifying the glycan structures that are present

Anti-Glycan Antibody Services

As stated above, the use of anti-glycan antibodies for characterization of cell-surface glycoconjugates has exhibited many advantages. However, the difficulty in the development of anti-glycan antibodies that target different glycan families and epitopes has hindered their applications. With innovative strategies and vast experience in the antibody development field, Creative Biolabs is now ready to assist our clients with their revolutionary projects in anti-glycan antibody development.

Our seasoned scientists can tailor custom-specific strategies to satisfy various research demands. If you are interested in our services, contact us to discuss your requirements.

Reference:

1. Oyelaran, O.; Gildersleeve, J. C. Glycan arrays: recent advances and future challenges. Current opinion in chemical biology. 2009, 13(4): 406-413.