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Development of Bioluminescent Cell-based Assay Platforms for Quantitative Measurement of ADCC/ADCP Activities for SARS-CoV-2 Antibodies

Part # PS401


Jun Wang, Pete Stecha, Jim Hartnett, Frank Fan, Mei Cong and Zhijie Jey Cheng
Promega Corporation

SARS-CoV-2 is a newly emerged coronavirus responsible for the current COVID19 pandemic. SARS-CoV-2 relies on its surface spike protein to bind to human host cell receptor angiotensin-converting enzyme 2 (ACE2) which is a step critical for viral entry, and thus spike protein has been the main target of antiviral mAb therapy and vaccine development. However, the mechanisms of anti-spike neutralizing antibodies are still not fully understood. Besides neutralization by blocking its interaction with ACE2, anti-spike antibodies may have additional antiviral activitties mediated by Fc domain, including antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). Therefore, it is very important to develop quantitative methods to help understand how anti-spike antibodies play protective and potential pathogenic roles to guide drug design and clinical development.

Here, we describe development of two bioluminescent cell-based assay platforms for quantitively measuring Fc-mediated effector functions for SARS-CoV-2 spike antibodies.:

  1. FcgR ADCC/ADCP Reporter Bioassays using engineered reporter effector cells and engineered target cells stably expressing SARS-CoV-2 Spike protein
  2. PBMC ADCC Assay using primary PBMC and engineered target cells stably expressing SARS-CoV-2 Spike protein and HaloTag-HiBiT