A Novel System for the Quantification of the ADCC Activity of Therapeutic Antibodies. (27th September 2017)

• Record Type: Journal Article
• Authors: Lallemand, Christophe; Liang, Feifei; Staub, Flore; Simansour, Maud; Vallette, Benoit; Huang, Lue; Ferrando-Miguel, Rosa; Tovey, Michael G.
• Other Names: Hooper Douglas C. Academic Editor.
• Is Part Of: Journal of immunology research. Volume 2017(2017)
• Publication Date: 2017-09-27
• Subjects: Immunology -- Periodicals; Immunology -- Research -- Periodicals; 616.07905
• DOI: 10.1155/2017/3908289
• Languages: English
• ISSNs: 2314-8861

Abstract

Abstract : Novel ADCC effector cells expressing the V-variant or F-variant of Fc γ RIIIa (CD16a) and firefly luciferase under the control of a chimeric promoter incorporating recognition sequences for the principal transcription factors involved in Fc γ RIIIa signal transduction, together with novel target cells overexpressing a constant high level of the specific antigen recognized by rituximab, trastuzumab, cetuximab, infliximab, adalimumab, or etanercept, confer improved sensitivity, specificity, and dynamic range in an ADCC assay relative to effector cells expressing a NFAT-regulated reporter gene and wild-type target cells. The effector cells also contain a normalization gene rendering ADCC assays independent of cell number or serum matrix effects. The novel effector and target cells in a frozen thaw-and-use format exhibit low vial-to-vial and lot-to-lot variation in their performance characteristics reflected by CVs of 10% or less. Homologous control target cells in which the specific target gene has been invalidated by genome editing providing an ideal control and a means of correcting for nonspecific effects were observed with certain samples of human serum. The novel effector cells and target cells expressing noncleavable membrane-bound TNF α have been used to quantify ADCC activity in serum from patients with Crohn's disease treated with infliximab and to relate ADCC activity to drug levels.