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

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• 作者：July Dorion-Thibaudeau^a ; ^b ; Yves Durocher^b ; ^c ; ¹ ; ^{yves.durocher@nrc-cnrc.gc.ca} ; Gregory De Crescenzo^a ; ¹ ; ^{gregory.decrescenzo@polymtl.ca}
• 关键词：FcγR ; Monoclonal antibody ; Surface plasmon resonance (SPR) ; Bioprocess monitoring
• 刊名：Molecular Immunology
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：82
• 期：Complete
• 页码：46-49
• 全文大小：691 K
• 卷排序：82

文摘

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 48 h 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.