Journal of Clinical and Cellular Immunology
Open Access
ISSN: 2155-9899
ISSN: 2155-9899
Michael G Tovey
Ecole Normale Sup�©rieure de Cachan, France
Posters & Accepted Abstracts: J Clin Cell Immunol
The activity of a number of monoclonal antibodies is mediated in part by antibody-dependent cell-mediated cytotoxicity (ADCC). Traditional methods for quantifying ADCC activity are labor intensive and have a high level of inherent variability. This is due in part to the use of primary human NK-cells from different donors as the effector cells and the use of a complex endpoint that is difficult to standardize, namely cytotoxicity. These limitations have been overcome by the use of an engineered effector cell line expressing the low affinity Fc receptor, Fc�³RIIIa (CD16) that responds to ligation of the Fc moiety of antibody bound to the specific antigen expressed on target cells by activation of a reporter gene. The use of novel engineered effector cells together with engineered target cells that over-express a constant high level of a specific antigen and the homologous control cells in which the gene encoding the specific antigen has been invalidated by genomic editing, provide the basis for the establishment of a precise and highly sensitive assay for the quantification of ADCC activity. Novel effector cells and engineered target B-cells that have been engineered to over-express a constant level of CD20,together with homologous control B-cells that do not express CD20, have been used to quantify the ADCC activity of Rituxan with a high degree of precision and sensitivity and with minimal interference from human serum.
Email: mtovey@ens-paris-saclay.fr