title: Stable expression of chimeric heavy chain antibodies in CHO cells.
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.
authors.affiliation: Biotechnology Research Institute, National Research Council of Canada, Montreal, QC, Canada.
authors.forename: Vishal
authors.initials: V
authors.lastname: Agrawal
authors.forename: Igor
authors.initials: I
authors.lastname: Slivac
authors.forename: Sylvie
authors.initials: S
authors.lastname: Perret
authors.forename: Louis
authors.initials: L
authors.lastname: Bisson
authors.forename: Gilles
authors.initials: G
authors.lastname: St-Laurent
authors.forename: Yanal
authors.initials: Y
authors.lastname: Murad
authors.forename: Jianbing
authors.initials: J
authors.lastname: Zhang
authors.forename: Yves
authors.initials: Y
authors.lastname: Durocher
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D011993:Recombinant Fusion Proteins