Primary Cell Bioassays
Primary peripheral blood mononuclear cells (PBMCs) are routinely used in traditional bioassays to quantify antibody drug potency in antibody-dependent cellular cytotoxicity (ADCC).
Promega primary cell bioassays provide a convenient bioluminescent detection method for target cell killing. These assays are biologically relevant, MOA-based assays that can be used to measure the potency and stability of antibodies and other biologics that specifically bind and activate Fcγ receptors. The assays include ADCC-qualified PBMCs and a choice of target cells stably expressing HiBiT fusion proteins. Upon lysis of the target cells and addition of detection reagent, a bright bioluminescent signal is generated.
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What are Primary Cell Bioassays?
Measurement of antibody-dependent cellular cytotoxicity (ADCC) is critical for understanding antibody Fc effector functions during monoclonal antibody development. Classic ADCC assays measure the short-term cytotoxicity of target cells typically preloaded with radioactive or fluorescent dyes after exposure to antibody-bound primary peripheral blood mononuclear cells (PBMCs) or the subset natural killer (NK) cells. Conventional ADCC assays are widely used in antibody discovery and characterization during early drug development, but the use of primary effector cells make them vulnerable to high assay variability due to high donor-to-donor variation. Therefore, these conventional assays are not suitable for use in a quality-controlled environment during product manufacture and development.
To improve ADCC assay consistency and robustness, Promega has partnered with BioIVT to develop a new PBMC ADCC bioassay with significant improvements to both PBMC effector cells and target cells. The new PBMC ADCC assay provides ADCC-qualified primary PBMCs that are produced by BioIVT and functionally QC tested in an ADCC assay by Promega. The assays provide a choice of engineered target cells stably expressing HiBiT proteins, instead of radioactive or fluorescent markers, to provide lower spontaneous release and better sensitivity. When HiBiT-expressing target cells are incubated with an antibody and PBMCs, the target cells are lysed and release HiBiT proteins. Subsequent binding of HiBiT to the complementary polypeptide LgBiT in the detection reagent constitutes a functional NanoLuc® Luciferase molecule to generate a bright luminescent signal. The bioassay is simple, homogeneous, highly sensitive and provides a robust assay window. It shows antibody potency comparable with the ADCC Reporter Bioassay in ADCC bridging studies.