title: Quantification and simultaneous evaluation of the bioactivity of antibody produced in CHO cell culture-The use of the ectodomain of FcγRI and surface plasmon resonance-based biosensor.
abstract: A surface plasmon resonance (SPR)-based assay has been developed in order to quantify the monoclonal antibody (Mab) Trastuzumab within the supernatant of a mammalian cell culture using the ectodomain of FcγRI (CD64) and confirm Mab bioactivity, i.e. binding to its antigen Her2, in a single biosensing experiment. Under partial mass transport limitation, we were able to quantify Mab present in unpurified samples taken throughout the cell culture. While Mab capture on the biosensor surface confirmed the ability of its Fc region to bind to FcγRI, the binding activity of its Fab region was also tested by injecting increasing concentrations of the Mab antigen (Her2). The kinetics of the interactions we recorded from 48h post transfection until the end of the culture, were superimposable, which highly suggested that the quality attributes of the antibody were conserved throughout the process. This SPR methodology is thus of great interest for atline quality control analysis during Mab production campaign.
authors.affiliation: Department of Chemical Engineering, Groupe de Recherche en Sciences et Technologies Biomédicales, Bio-P2 Research Unit, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montreal, QC H3C 3A7, Canada; Life Sciences, NRC Human Health Therapeutics Portfolio, Building Montreal-Royalmount, National Research Council Canada, Montreal QC H4P 2R2, Canada.
authors.forename: July
authors.initials: J
authors.lastname: Dorion-Thibaudeau
authors.affiliation: Life Sciences, NRC Human Health Therapeutics Portfolio, Building Montreal-Royalmount, National Research Council Canada, Montreal QC H4P 2R2, Canada; Biochemical Department, Université de Montréal, Montreal, QC H3C 3J7, Canada. Electronic address: yves.durocher@nrc-cnrc.gc.ca.
authors.forename: Yves
authors.initials: Y
authors.lastname: Durocher
authors.affiliation: Department of Chemical Engineering, Groupe de Recherche en Sciences et Technologies Biomédicales, Bio-P2 Research Unit, École Polytechnique de Montréal, P.O. Box 6079, succ. Centre-Ville, Montreal, QC H3C 3A7, Canada. Electronic address: gregory.decrescenzo@polymtl.ca.
authors.forename: Gregory
authors.initials: G
authors.lastname: De Crescenzo
mesh_terms: D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D017452:Receptors, IgG; D020349:Surface Plasmon Resonance / Q000379:methods*; D000068878:Trastuzumab / Q000032:analysis*
chemical_list: D017452:Receptors, IgG; D000068878:Trastuzumab
keywords: Bioprocess monitoring; FcγR; Monoclonal antibody; Surface plasmon resonance (SPR)