Stable expression of chimeric heavy chain antibodies in CHO cells

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DOIResolve DOI: http://doi.org/10.1007/978-1-61779-968-6-18
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TypeArticle
Journal titleMethods in Molecular Biology
ISSN1064-3745
ISBN9781617799679
Volume911
Pages287303; # of pages: 17
Subjectantibiotic g 418; blasticidin S; chimeric antibody; chimeric heavy chain antibody; hygromycin B; phleomycin; plasmid vector; polyethyleneimine; puromycin; unclassified drug; hybrid protein; immunoglobulin heavy chain; monoclonal antibody; animal cell; antibody production; article; CHO cell; cloning; controlled study; culture technique; drug activity; genetic transfection; heavy chain; nonhuman; priority journal; protein expression; suspension cell culture; toxicity; animal; gene expression; gene order; gene vector; genetics; hamster; isolation and purification; metabolism; Animals; Antibodies, Monoclonal; CHO Cells; Cricetinae; Gene Expression; Gene Order; Genetic Vectors; Immunoglobulin Heavy Chains; Recombinant Fusion Proteins; Transfection
AbstractCamelid 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.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Biotechnology Research Institute (BRI-IRB); NRC Institute for Biological Sciences (IBS-ISB)
Peer reviewedYes
NPARC number21269570
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Record identifier3b08d77b-835e-464b-8195-def3eef78cf8
Record created2013-12-12
Record modified2016-05-09
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