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ADCs, mAbs, CAR Ts: One Bioluminescent Platform Measures Them All

The HiBiT Target Cell Killing (TCK) platform, with off-the-shelf, mix-and-match components, measures four killing mechanisms for broad immunotherapy modality assessment

September 8, 2026

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A crucial step during immunotherapy development is verifying MoA through delivery of the cytotoxic payloads to the targeted cells in a heterogeneous environment. The streamlined Promega HiBiT Target Cell Killing (TCK) platform is a bioluminescent cell-based system that measures cytotoxicity with specificity, simplicity, and sensitivity during therapeutic development. The mix-and-match platform supports broad immunotherapy modality assessment for four killing mechanisms—CAR-T cell-mediated killing, antibody-dependent cellular cytotoxicity (ADCC), T cell-dependent cellular cytotoxicity (TDCC), and antibody-dependent cellular phagocytosis (ADCP).

Off-the-shelf, thaw-and-use HiBiT target cells express an intracellular HiBiT peptide that remains stable in media for >3 days with minimal leakage and is released only upon cell death. Released HiBiT binds cell-impermeable LgBiT to form functional NanoBiT luciferase, generating a bright, quantitative signal. The luminescent signal is proportional to target cell death alone, with no contribution from effector cells, making the platform ideal for co-culture experiments.

Thaw-and-use target cells

A growing library of off-the-shelf assay components includes thaw-and-use, functionally tested cell lines that address blood cancer targets, including B cell lymphoma and leukemia, myeloid leukemia, and multiple myeloma (Raji, Ramos, H929), as well as solid tumor targets for ovarian carcinoma (OVCAR3), breast adenocarcinoma, and lung carcinoma (SKOV3, SK-BR-3). HiBiT-containing target cell lines can also be custom tailored using ViaScript^® transfection.

Principle of the HiBiT TCK Bioassay. Cytotoxic mAbs and/or effector cells are incubated with target cells expressing a HiBiT fusion protein. Upon killing of the target cell, the HiBiT fusion protein is released and binds extracellular LgBiT to create a functional NanoBiT^® Luciferase enzyme. Luminescence is measured using a luciferase substrate and the GloMax^® Discover System.

For cell lines not in the standard panel, the ViaScript (HiBiT) TCK Bioassay enables rapid screening of target cells using transient HiBiT expression. ViaScript is a novel mRNA transfection reagent that enables rapid and titratable transient expression of HiBiT mRNA in a wide range of adherent or suspension target cell types for early high-throughput screening and clone selection. The transfected cell lines can be screened simultaneously and used in a HiBiT TCK Bioassay and paired with qualified primary effector cells and a biologic to measure ADCC, ADCP, TDCC or CAR-T cell killing. Once optimal TCK lines are identified with the ViaScript (HiBiT) TCK Bioassay, Promega’s Tailored R&D Solutions (TRS) offers custom clonal line development.

Off-the-shelf primary effector cells

The homogeneous, sensitive HiBiT TCK platform and bioassay provides a robust assay window for four immunotherapy killing mechanisms. The versatile mix-and-match platform supports experiments with different combinations of effector and target cells. The resulting luminescent signal is specific to target cell killing, making the bioassay well-suited for mixed co-culture experiments during development efforts. The HiBiT TCK platform supports ADCC with PBMC effectors, TDCC with CD8+ T cells, ADCP with macrophages, and CAR-T killing assays across a 4–72-hour time course.

Explore More promega.com/products/reporter-bioassays/target-cell-killing-bioassays.