A Plate Based Real-Time Annexin V Method for Monitoring Antibody Drug Conjugate Induced Apoptosis and Cell Death

Part # PS314


Steven Edenson1, Kevin Kupcho1, Andrew Niles1, John Shultz1, Jamison Grailer1, Wenhui Zhou2, Robin Hurst1, Jim Hartnett1, Terry Riss1, Dan Lazar1, and James Cali1
1Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711
2Promega Biosciences LLC, 277 Granada Drive, San Luis Obispo, CA 93401

Antibody drug conjugates (ADC) induce cell death via cognate interactions with surface antigens, internalization, release of toxin, and the binding of the toxin to its cellular targets. The key factors that determine the effectiveness of an ADC include the antibody, linker, toxin, and conjugation method. Testing all possible combinations can be challenging with assays that simply readout a single time point. To address this need, we developed a real-time, live cell assay method that utilizes a fully homogeneous, bioluminescent annexin V reagent. The method does not require laborious washing and sample preparation steps associated with traditional annexin methods and is fully compatible with plate-based multimodal signal detection systems. This work describes our efforts to characterize and compare the performance of the bioluminescent annexin assay to traditional endpoint assays after ADC exposure with target cells.

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