title: Towards the development of a surface plasmon resonance assay to evaluate the glycosylation pattern of monoclonal antibodies using the extracellular domains of CD16a and CD64.
abstract: We here report the production and purification of the extracellular domains of two Fcγ receptors, namely CD16a and CD64, by transient transfection in mammalian cells. The use of these two receptor ectodomains for the development of quantitative assays aiming at controlling the quality of monoclonal antibody production lots is then discussed. More specifically, the development of surface plasmon resonance-based biosensor assays for the evaluation of the glycosylation pattern and the aggregation state of monoclonal antibodies is presented. Our biosensor approach allows discriminating between antibodies harboring different galactosylation profiles as well as to detect low levels (i.e., less than 2%) of monoclonal antibody aggregates.
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; Biochemistry Department, Université de Montréal, Montreal, QC H3C 3J7, Canada.
authors.forename: Céline
authors.initials: C
authors.lastname: Raymond
authors.affiliation: Chemistry Department, University of Manitoba, 144 Dysart Road, Winnipeg, MB R3T 2N2, Canada.
authors.forename: Erika
authors.initials: E
authors.lastname: Lattová
authors.affiliation: Chemistry Department, University of Manitoba, 144 Dysart Road, Winnipeg, MB R3T 2N2, Canada.
authors.forename: Helene
authors.initials: H
authors.lastname: Perreault
authors.affiliation: Life Sciences, NRC Human Health Therapeutics Portfolio, Building Montreal-Royalmount, National Research Council Canada, Montreal, QC H4P 2R2, Canada; Biochemistry 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; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D015374:Biosensing Techniques; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D051916:Membrane Cofactor Protein / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D017452:Receptors, IgG / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D020349:Surface Plasmon Resonance; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; C497857:CD46 protein, human; C496124:FCGR3A protein, human; D051916:Membrane Cofactor Protein; D017452:Receptors, IgG
keywords: Aggregation; Anti-histidine capture; CD16a; CD64; Glycosylation; Monoclonal antibody; Surface plasmon resonance (SPR)