title: An efficient detection agent for the high throughput screening of recombinant manufacturing cell lines.
abstract: To ensure the selection of high producing recombinant cell lines, a number of screening processes were developed in the presence of detection agents. Here, CHO cell lines secreting recombinant antibodies were detected in semi-solid medium containing detection agents. The aim was to compare two protein A-derived detection agents to two commercial fluorescent antibodies directed against the Fc part of the antibody of interest: the protein A derived Z domain fused to the red fluorescent protein and protein A labelled with a fluorescent Dylight™ 488 dye. All of these agents were compatible with cell recovery and colony formation, and specifically detected colonies secreting recombinant antibodies. Optimisation of the concentration of the fluorescent protein A allowed the identification of a higher number of good producers. Thus these data demonstrate that fluorescently labelled protein A-derivatives can be used for the selection of high producer cells.
authors.affiliation: Cell Sciences, MerckSerono SA, ZI B, CH-1809 Fenil-sur-Corsier, Switzerland. Electronic address: Valerie.Duverger@merckgroup.com.
authors.forename: Valérie
authors.initials: V
authors.lastname: Duverger
authors.forename: Christophe
authors.initials: C
authors.lastname: Sauvage
authors.forename: Michel
authors.initials: M
authors.lastname: Kobr
authors.forename: Markus O
authors.initials: MO
authors.lastname: Imhof
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D018929:Cell Culture Techniques; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000379:methods*; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods*; D013205:Staphylococcal Protein A / Q000032:analysis* / Q000276:immunology; D019076:Transgenes / Q000235:genetics; D000097573:Red Fluorescent Protein
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D000097573:Red Fluorescent Protein; C048844:anti-IgG
keywords: FU; Fluorescently labelled protein A; GSEM; High throughput; MSX; Manufacturing recombinant cell line; fluorescence unit.; glutamine synthetase expression medium supplement; l-methionine sulfoximine