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Application: Luciferase- based analysis of live and dead target cells in cytotoxicity assays.Test the efficacy of multi-specific immune engager molecules.Assess the Fc effector function of candidate antibodies.
Background: Lymphocyte-mediated cytotoxicity is a form of cellular immunity against intracellular pathogens, including viruses and certain bacteria and parasites. The most popular in vitro methods to monitor lymphocyte-mediated cytotoxicity on target cells are cell-mediated cytotoxicity assays such as ADCC (antibody-dependent cellular cytotoxicity) and TDCC (T-cell dependent cellular cytotoxicity) in which immune effector cells and target cells are co-cultured. To analyze immune effector cell cytolytic activity in such heterogeneous cell population of effector and target cells, it is important to be able to discriminate between effector and target cell populations with distinct phenotypes. The use of luciferase allows for a clear separation between the effector and the surviving target cells. The instability of firefly luciferase when released from dead target cells in cell culture gives it a half-life of approximately 2 hours, eliminating any residual luminescence signal generated from dead target cells. Cytotoxicity assays are crucial to understand the potency of CAR (chimeric antigen receptor) T and NK cells, and antibody-based immunotherapies.
Description: The PBMC Cytotoxicity Luciferase Assay Kit (OVCAR3) is a kit designed to determine the cytotoxicity profile of PBMCs (Peripheral Blood Mononuclear Cells) towards the eGFP/Firefly Luciferase OVCAR3 Cell Line. It uses the luminescence signal from eGFP/Firefly Luciferase OVCAR3 Cell Line to measure the number of live target cells within a mixed cell population of PBMC and eGFP/Firefly Luciferase OVCAR3 cells. The kit contains PBMCs, eGFP/Firefly Luciferase OVCAR3 Cell Line, cell culture media and One-Step™ Luciferase Assay System. In addition, this kit includes a positive control bispecific antibody.
Host Cell Line: OVCAR3, ovarian adenocarcinoma cell line, adhering
Storage Stability: Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage. Contact technical support at support@bpsbioscience.com if the cells are not frozen in dry ice upon arrival.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: BSL-1
References: Bradbury A., et al., 2020 Cancers (Basel) 12(7):1939.
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