Development of a Bioluminescent Cell-Based Assay to ... - Promega

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ADCC Reporter Bioassay demonstrates appropriate specificity Target cells, effector cells and specific antibody Confidential and Proprietary. Not for Further Disclosure. No Target cells No Effector cells or no FcgRIIIa No antibody or non-specific antibody Assay signal is dependent on: Presence of Target cells + Presence of FcgRIIIa receptor + Appropriate specific antibody 8
Bioassay development: Optimizing Effector:Target (E:T) ratio An example: Evaluation of different E:T ratios for assay optimization using frozen, thaw-and-use engineered Jurkat Effector cells (FcgRIIIA (V158) / NFAT-RE-luc2) with fresh-from-culture target cells (WIL2-S, CD20+). • Fixed Effector cell number • Varied Target cell number and E:T ratios • 6hr induction • Bio-Glo Assay Reagent Confidential and Proprietary. Not for Further Disclosure. Higher target cell numbers generate greater fold induction 9
Page 1 and 2: Development of a Bioluminescent Cel
Page 3 and 4: Signaling pathways activated in eff
Page 5 and 6: Protocol 1. Reference or test antib
Page 7: FcgRIIIa antibody blocking of ADCC
Page 11 and 12: Bioassay development: Evaluating cl
Page 13 and 14: Target cells can also be frozen, th
Page 15 and 16: ADCC Reporter Bioassay using frozen
Page 17 and 18: Precision, accuracy and linearity o
Page 19 and 20: ADCC Reporter Bioassay is stability
Page 21 and 22: Additional target cell option: Raji
Page 23 and 24: iological activity of therapeutic a
Page 25 and 26: Planned ADCC Reporter Bioassay kit
Page 27: For more information Confidential a

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