Target Cell Killing Bioassays

Our HiBiT Target Cell Killing (TCK) Bioassays provide a simple, sensitive and highly specific method to measure target cell killing induced by a variety of biologic drugs. Upon lysis, the assays generate a bright luminescent signal that can be measured using a standard luminometer.

How Do TCK Bioassays Work?

Faster Processing Times, Dynamic Signal-to-Noise Ratio and Easy-to-Interpret Data

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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 Microplate Reader.

TCK Cell Lines for Immuno-Oncology Research

Primary Effector Cells


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PBMCs
  • ADCC-qualified
  • Single-donor derived
  • FcγR genotyped
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CD8+ T cells
  • TDCC qualified
  • Functionally tested
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Macrophage
  • ADCP qualified
  • Functionally tested
  • FcγR genotyped

Solid Tumor Target Cells


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Ovarian Carcinoma
  • OVCAR3 and SKOV3
  • Expressing: HER2, MSLN, 5T4, MUC16
  • MSLN-KO line available
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Breast Adenocarcinoma
  • SK-BR-3
  • Expressing: HER2, EpCAM
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Lung Carcinoma
  • A549
  • Expressing: EGFR
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Melanoma
  • A375
  • Expressing: HER2, CD70, B7-H3

Blood Cancer Target Cells


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B cell Lymphoma and Leukemia
  • Raji
  • Expressing: CD19, CD20, CD22, CD38
  • CD19, CD20 and CD19/CD20 KO lines available
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B cell Lymphoma and Leukemia
  • Ramos
  • Expressing: CD19,CD20, CD22, CD38
  • CD19 KO line available
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Myeloid Leukemia
  • U937 and K562
  • Expressing: CD33 and CLL-1
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Multiple Myeloma
  • H929
  • Expressing: BCMA and CD38
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T cell Leukemia
  • T2
  • Expressing: CD5, CD7, CD30, and CD52

Images created with BioRender.com

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Applications

The HiBiT TCK platform can be used to measure TCK activity induced by a variety of biologic drug modalities. The data included here
are representative of three applications: CAR-T cell-mediated cytotoxicity, antibody-dependent cell-mediated cytotoxicity (ADCC), and
T cell-dependent cell-mediated cytotoxicity (TDCC).

CAR-T Cell-Mediated Cytotoxicity

Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)

T Cell-Dependent Cell-Mediated Cytotoxicity (TDCC)

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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.

TCK activity measured using the PBMC ADCC Bioassay. PBMC-mediated killing of Raji (HiBiT) Target Cells by anticancer therapeutic mAb, rituximab.

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.)