1. Stable expression of chimeric heavy chain antibodies in CHO cells
- Author
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Agrawal, V., Slivac, I., Perret, S., Bisson, L., St-Laurent, G., Murad, Y., Zhang, J., and Durocher, Y.
- Subjects
Recombinant Fusion Proteins ,Genetic Vectors ,puromycin ,cloning ,gene order ,plasmid vector ,Gene Expression ,animal cell ,CHO Cells ,Transfection ,hygromycin B ,drug activity ,Cricetinae ,gene vector ,chimeric heavy chain antibody ,blasticidin S ,genetics ,suspension cell culture ,heavy chain ,protein expression ,hybrid protein ,antibody production ,CHO cell ,isolation and purification ,phleomycin ,toxicity ,Antibodies, Monoclonal ,immunoglobulin heavy chain ,chimeric antibody ,unclassified drug ,culture technique ,hamster ,antibiotic g 418 ,monoclonal antibody ,genetic transfection ,polyethyleneimine ,Immunoglobulin Heavy Chains ,metabolism - Abstract
Camelid single domain antibodies fused to noncamelid Fc regions, also called chimeric heavy chain antibodies (cHCAb), offer great potential as therapeutic and diagnostic candidates due to their relatively small size (80 kDa) and intact Fc. In this chapter, we describe two approaches, limiting dilution and minipools, for generating nonamplified Chinese hamster ovary cell lines stably expressing cHCAb in suspension and serum-free cultures using a stringent antibiotic selection. Neither of the protocols necessitates the acquisition or implementation of expensive automated infrastructures and thus could be applied in any lab with minimal cell culture setup. The given protocol allows the isolation of stable clones capable of generating up to 100 mg/L of antibody in batch mode performed in shaker flasks. © 2012 Springer Science+Business Media, LLC.
- Published
- 2012
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