Application Note


Easy to use, precise and scalable bioluminescent ADCC Reporter Bioassay that correlates well to classic ADCC bioassay


By Terry Surowy, Zhi-Jie (Jey) Cheng, Rich Moravec, Aileen Paguio, Denise Garvin, Neal Cosby and Frank Fan, Promega Corporation


I


n recent years the pharmaceutical industry has been focusing more on biologics to fill its R&D pipelines or in many cases looking to develop biosimilars or biobetters for marketed drugs. Monoclonal Ab drug development leads the pack of biologics, accounting for one- third of medicines in development1.


much more complex to develop. Because of their complex structure and complicated manufacturing processes, regulatory agen- cies are looking for better functional data to support new drug claims to ensure effi- cacy and safety.


Although biologics promise high therapeu- tic index and potential reduced toxicity compared to small molecule drugs, they are


The European Medicines Agency (EMA) and the US Food and Drug Administration (FDA) have each drafted guidelines2,3 for biosimilars product development. Both rec- ommend extensive structural and function- al characterisation of the proposed biother- apeutic including bioassay data reflective of the drug’s mechanism of action (MOA). Such functional assays include antibody- dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxi- city (CDC) and complement activation to address functional aspects of the mAb. Classic ADCC bioassays are well-known for tedious preparation of primary effector cells and assay variability. In contrast, the novel ADCC Reporter Bioassay is simple to use and more precise4. Here we demonstrate how data generated with the novel ADCC Reporter Bioassay correlates well to that obtained using classic ADCC bioassay based on LDH release. Additionally, we demon- strate the assay is easily converted to 384- well format. These data enable broader application of the ADCC bioassay to mono- clonal Ab drug discovery and development, and should help provide better functional data for regulatory agency review.


Figure 1: Principle of the ADCC Reporter Bioassay. Antibody binds target cell antigens and FcRIIIa receptors expressed on effector cells, just as in classic ADCC assays. In the ADCC Reporter Bioassay, luminescent signal is generated following specific antibody binding to target and engineered, Jurkat- derived effector cells and consequent induction of the NFAT pathway in the engineered effector cells


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ADCC Reporter Bioassay Principle


The ADCC Reporter Bioassay (www.promega.com) uses engineered


Drug Discovery World Spring 2013


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