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

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Evaluating passage and cell density at freezing for WIL2-S (CD20+) Target cells Establishment of optimized frozen, thaw-and-use Target cells includes establishment of acceptable passage range and cell growth conditions prior to freezing. Important for establishing cell banks and manufacturing capabilities Confidential and Proprietary. Not for Further Disclosure. Fold induction 30 25 20 15 10 5 EC50 Pass 8 Pass 12 Pass 18 Pass 24 0 0.01 0.1 1 10 100 1000 Final Rituximab (ng/ml) Pass 8 8.851 Pass 12 7.057 6 hr assay Effector:Target ratio = 6 Pass 18 6.520 Pass 24 7.122 14
ADCC Reporter Bioassay using frozen, thaw-and-use Effector and Target cells A. Fresh cells from continuous culture B. Frozen, thaw-and-use cells Shown are representative data from ADCC reporter assay using fresh (A), or frozen thaw-and-use (B) Jurkat effector cells and Wil2-S cells. Target cells are CD20+ WIL2-S EC50s and fold induction of responses are equivalent Confidential and Proprietary. Not for Further Disclosure. 15
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 and 8: FcgRIIIa antibody blocking of ADCC
Page 9 and 10: Bioassay development: Optimizing Ef
Page 11 and 12: Bioassay development: Evaluating cl
Page 13: Target cells can also be frozen, th
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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