HiBiT Target Cell Killing Platform
The HiBiT target cell killing (TCK) bioassay platform enables highly sensitive and specific measurement of target cell killing induced by a variety of biologic drugs, including monoclonal antibodies (mAbs), bispecific antibodies and CAR-T cells. The assay is simple and homogenous. Upon target cell lysis, the assay produces a bright luminescent signal that can be measured using a standard luminometer.
- Simple, fast, and sensitive technology to specifically measure killing of target cells
- Flexible platform to measure the activity of a variety of biologic drugs
- Off-the-shelf HiBiT Target Cells expressing common immunotherapy targets (CD19, CD20, BCMA, and more)
- Custom development of HiBiT Target Cells and assay optimization to meet your needs
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.
Available HiBiT Target Cells
|Target Cells||Cell Line Background||Drug Targets||Potential Applications||Test Drugs Available*|
|A549||Lung carcinoma||EGFR||mAb, Bispecific, CAR-T||cetuximab, trastuzumab, daratumumab (ADCC)|
|BCMA K562||Chronic myelogenous leukemia||BCMA||ADC, Bispecific, CAR-T||
|CD19 K562||Chronic myelogenous leukemia||CD19||mAb, Bispecific, CAR-T||blinatumomab (TDCC)|
|CIITA K562||Chronic myelogenous leukemia||N/A||mAb, Bispecific, CAR-T||blinatumomab (TDCC)|
|Claudin 18.2 CHO-K1||Chinese hamster ovarian||Claudin 18.2||mAb, Bispecific, CAR-T||zolbetuximab (ADCC)|
|H929||Multiple myeloma||BCMA, CD38||ADC, Bispecific, CAR-T||
|K562||Chronic myelogenous leukemia||N/A (Parental)||mAb, Bispecific, CAR-T||—|
|Membrane TNFα CHO-K1||Chinese hamster ovarian||TNFα||mAb||adalimumab, golimumab, certolizumab (ADCC)|
|OVCAR3||Ovarian carcinoma||MSLN, 5T4, WT, HER2||mAb, Bispecific, CAR-T||ertumaxomab (TDCC)|
|Raji (also CD19-, CD20- & CD19-/CD20- control cell lines)||B lymphoma||CD19, CD20, CD22, CD38||mAb, ADC, Bispecific, CAR-T||
rituximab, daratumumab (ADCC)
|Ramos (also CD19- control cell line)||B lymphoma||CD19, CD20, CD22, CD38||mAb, ADC, Bispecific, CAR-T||
rituximab, daratumumab (ADCC)
|SARS-CoV-2-S CHO-K1||Chinese hamster ovarian||SARS-CoV-2 S Protein||mAb, Bispecific, CAR-T||—|
|SK-BR-3||Breast cancer||HER2, EpCAM||mAb, Bispecific, CAR-T||trastuzumab (ADCC)|
|SKOV3||Ovarian carcinoma||MSLN, 5T4, MUC16, HER2, PD-L1||mAb, ADC, Bispecific, CAR-T||
|T2||Lymphoma||HLA-A2+ for TCR-T||TCR-T||—|
|U937||Lymphoma||CD33, CLL-1||mAb, Bispecific, CAR-T||gemtuzumab (ADC)|
*Drugs are commercially available for the following targets: blinatumomab: CD3xCD19 BiTE; rituximab: anti-CD20; daratumumab: anti-CD38; cetuximab: anti-EGFR; trastuzumab: anti-HER2.
CAR-T Cell-Mediated Cytotoxicity
Extended assay incubation time reveals serial TCK activity. T cells transduced with CAR-19 or a GFP control lentivirus were combined with HiBiT Target cells (Ramos) at different effector:target (E:T) ratios. After incubation for 24 hours (Panel A) or 48 hours (Panel B), NanoBiT® Extracellular Detection Reagent was added, and luminescence was read on a GloMax® Discover plate reader. The EC50 shifts left over time, indicating serial TCK at lower E:T ratios.
Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)
TCK activity measured using the PBMC ADCC Bioassay. Panel A. PBMC-mediated killing of Raji (HiBiT) Target Cells by anticancer therapeutic mAb, rituximab. Panel B. PBMC-mediated killing of SARS-CoV-2 S CHO-K1 (HiBiT) target cells by an anti-SARS-CoV-2 spike (S2) antibody.
T Cell-Dependent Cell-Mediated Cytotoxicity (TDCC)
The TDCC Assay measures target cell-specific killing. Thaw and Use CD8+ T Cells (TDCC Qualified) were combined with HiBiT-expressing target cells and appropriate Bispecific T Cell Engagers (BiTEs.) After 18–24 hours incubation, NanoBiT® Extracellular Detection Reagent was added and luminescence was read on a GloMax® Discover plate reader. The assay enables sensitive measurement of bispecific antibody potency in a homogenous format with no medium transfer required.