The impact of the glycan distribution on the in vivo function and half-life of monoclonal antibodies has long motivated the genetic engineering of producer cells to achieve structures that enhance efficacy, safety and stability. To facilitate glycoengineering of IgG-producing Chinese hamster ovary cells, we present a rapid protocol that involves the use of RNA interference for the knockdown of genes of interest coupled with capillary gel electrophoresis and laser-induced fluorescence detection (CGE-LIF) for fast, high-throughput glycan analysis. We apply this methodology to the Fut8 gene, responsible for the addition of core fucose, which is a typical target for increasing antibody-dependent cellular cytotoxicity.
CITATION STYLE
Kotidis, P., Marbiah, M., Donini, R., Gómez, I. A., del Val, I. J., Haslam, S. M., … Kontoravdi, C. (2022). Rapid Antibody Glycoengineering in CHO Cells Via RNA Interference and CGE-LIF N-Glycomics. In Methods in Molecular Biology (Vol. 2370, pp. 147–167). Humana Press Inc. https://doi.org/10.1007/978-1-0716-1685-7_7
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