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.
Vishal Agrawal and Igor Slivac contributed equally to this chapter.
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The authors declare no competing interests. This is a NRC publication number.
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Agrawal, V. et al. (2012). Stable Expression of Chimeric Heavy Chain Antibodies in CHO Cells. In: Saerens, D., Muyldermans, S. (eds) Single Domain Antibodies. Methods in Molecular Biology, vol 911. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-968-6_18
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DOI: https://doi.org/10.1007/978-1-61779-968-6_18
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-61779-968-6
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