Rapid progress in understanding glycosylation pathways of eukaryotes came with the application of genetic strategies to isolate mutants of mammalian cells and yeasts with defects in glycans synthesis. Virtually any agent that recognizes cell-surface glycans or a specific surface glycoprotein, GPI-anchored protein, or glycolipid can be used to isolate mutants with a glycosylation defect. Here, Creative Biolabs gives an introduction to animal cell glycosylation mutants and the use of anti-glycan antibodies for selecting or isolating cell lines that express altered cell-surface glycans.
Usually, the selection or enrichment of animal cell glycosylation mutants is needed to find rare recessive of dominant mutants bearing the desired glycosylation phenotype. This can be achieved by using glycan-binding proteins or other agents (e.g., cytotoxic lectins) that recognize cell-surface glycans. Thus, anti-glycan antibodies to specific glycan determinants can be used as well. By using fluorescently tagged antibodies, mutants with glycosylation defects (either deficient in binding or with acquired novel binding ability because of altered glycosylation) can be enriched.
Fig.1 Cell glycosylation mutants selection. (Esko, 2017)
Glycosylation mutants of cultured animal cells can be used to address many questions in glycobiology and for glycosylation engineering of recombinant glycoproteins. These mutants generate glycans with truncated or altered structures, providing an opportunity to study functional roles for cell-surface glycans in the context of a living cell. Moreover, Chinese hamster ovary (CHO) mutants have been used in the production of recombinant therapeutic glycoproteins and glycosylation engineering. For example, LEC11 CHO cells have been used to generate proteins carrying sialyl LewisX.
The use of anti-glycan antibodies for selection of animal cell glycosylation mutants is highly specific and sensitive. However, there are limited commercial resources of anti-glycan antibodies targeting different glycan families and epitopes. Based on our sophisticated technologies and abundant experience, Creative Biolabs is now providing professional anti-glycan antibody development services to facilitate related research.
Our seasoned scientists are willing to communicate with our clients and design precisely tailored services to meet all kinds of expectations. If you are interested in our services, contact us to discuss your requirements.
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