Toward a Better Understanding of Bioassays for the Development of Biopharmaceuticals by Exploring the Structure-Antibody-Dependent Cellular Cytotoxicity Relationship in Human Primary Cells

• DOI: 10.3389/fimmu.2020.552596
• Publication Date: 2020-10-29T09:00:50Z
• Authors: Sébastien Wieckowski; Cécile Avenal; Arturo V. Orjalo; Daniel Gygax; Florian Cymer
• Keywords: natural killer cells; glycosylation; flow cytometry; Immunology; Antibody Affinity; Antibody-Dependent Cell Cytotoxicity; RC581-607; Trastuzumab; 3. Good health; trastuzumab; Structure-Activity Relationship; 03 medical and health sciences; breast cancer; FcγRIIIA; 0302 clinical medicine; Humans; Lymphocytes; Immunologic diseases. Allergy

Pharmaceutical manufacturing relies on rigorous methods of quality control drugs and in particular the physico-chemical functional characterizations monoclonal antibodies. To that end, robust bioassays are very often limited to reporter gene assays use immortalized cell lines supposed mimic immune cells such as natural killer (NK) detriment primary materials, which appreciated for their biological validity but also difficult exploit due great diversity between individuals. Here, we characterized phenotype peripheral blood circulating cytotoxic 30 healthy donors, repertoire markers, using flow cytometry. In parallel, antibody-dependent cellular cytotoxicity (ADCC) effector functions these by measuring cytolytic activity against a cancer cell-line expressing HER2 presence trastuzumab with regards FCGR3A genotype. We could not establish correlation or grouping individuals data generated from whole mononuclear cells, however isolation CD56-positive population, is composed only NK T (NKT) γδ-T well subsets activated monocytes dendritic made it possible standardize parameters ADCC enhance overall avidity without eliminating inter-individual diversity. Finally, CD56+ experiments comparing glycoengineered variants was conclusive test limits this type ex vivo system. Although reflected some extent vitro receptor binding properties NK92 previously published, reaching plateau limit framework.