WIL2-S and an Engineered Jurkat Cell Line as a Model for Assessment of Apoptotic and Phagocytic Activity Upon Treatment with Anti-CD20
Sinosha Paralikar, Keyuri Mokashi, Sheetal Raut and Sanjeev Gupta
Advanced Biotech Lab, Ipca Laboratories Ltd., India
*Corresponding Author: Sanjeev Gupta, Advanced Biotech Lab, Ipca Laboratories Ltd., India.
March 14, 2023; Published: April 11, 2023
I-RmAb is an anti-CD20 monoclonal antibody, that we have developed as a therapeutic to treat B-cell malignancies. It induces cell death via various mechanisms such as complement dependent cytotoxicity (CDC), antibody dependent cell mediated cytotoxicity (ADCC), antibody dependent cell mediated phagocytosis (ADCP), apoptosis and sensitization of cancer cells to immunotherapy.
While other in-vitro assays are well-established, in this research, we focus on comparing Fab mediated apoptosis induced by I-RmAb in two CD20 positive human lymphoblast cell lines - WIL2-S and Raji, using flow cytometry, thus identifying a time window of peak apoptosis which has not been explored in depth before. This method is difficult to develop because I-RmAb elicits a very low apoptotic response and varies within different cell lines and patient derived cells such as B-CLL. We have also explored a luminescence and fluorescence method, and concluded that flow cytometry is a more reliable technique to assess apoptosis. Through this study, we hope to further narrow down the exact time window of apoptotic activity and a cell line sensitive enough to detect the same for developing a robust assay.
Alongside, we also outline a developed method that demonstrates I-RmAb’s activation of the phagocytic pathway as a function of the FcγRIIa receptor in commercially available Jurkat cells harboring the NFAT reporter. Both apoptosis and ADCP assays are an important quality aspect for biosimilarity assessment with limited availability of experimental data in the literature. Hence, research under this wing will help improve the process of designing functional assays.
Keywords: Apoptosis; ADCP; Anti-CD20; WIL2-S
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