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

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

PBMCs

ADCC-qualified
Single-donor derived
FcγR genotyped

CD8+ T cells

TDCC qualified
Functionally tested

Macrophage

ADCP qualified
Functionally tested
FcγR genotyped

TCK Cell Line Screening

ViaScript® (HiBiT) TCK Bioassay

Transient expression of HiBiT
Identify optimal TCK line
For blood cancer or solid tumor cell lines

Solid Tumor Target Cells

Ovarian Carcinoma

OVCAR3 and SKOV3
Expressing: HER2, MSLN, 5T4, MUC16
MSLN-KO line available

Breast Adenocarcinoma

SK-BR-3
Expressing: HER2, EpCAM

Lung Carcinoma

A549
Expressing: EGFR

Melanoma

A375
Expressing: HER2, CD70, B7-H3

Blood Cancer Target Cells

B cell Lymphoma and Leukemia

Raji (HaloTag-HiBiT)
Raji (LDH-HiBiT)*
Ramos
Cell lines expressing CD19, CD20, CD22 and CD38

Knockout lines available for some targets

Myeloid Leukemia

U937 and K562
Expressing: CD33 and CLL-1

Multiple Myeloma

H929
Expressing: BCMA and CD38

T cell Leukemia

T2
Expressing: CD5, CD7, CD30, and CD52

*No functional differences have been observed between LDH and HaloTag® cell lines.

Images created with BioRender.com

Have you identified an optimal cell line with ViaScript® (HiBiT) TCK Bioassay?

Our Tailored R&D Solutions (TRS) Team builds clonal TCK lines.

Contact Us

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),
T cell-dependent cell-mediated cytotoxicity (TDCC) and antibody-dependent cellular phagocytosis (ADCP).

CAR-T Cell-Mediated Cytotoxicity

Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)

T Cell-Dependent Cell-Mediated Cytotoxicity (TDCC)

Antibody-Dependent Cellular Phagocytosis

Dose-response curve showing the percentage of antibody-dependent cellular phagocytosis (ADCP) as a function of trastuzumab concentration (log₁₀[g/ml]). The curve demonstrates a sigmoidal increase in ADCP, with maximal activity plateauing at higher concentrations.

CAR-T activity demonstrated using HiBiT Target Cells. 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 and incubated for 24 hours.

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.)

The Macrophage ADCP Bioassay quantifies phagocytic killing of target cells. Thaw and Use, ADCP-qualified human macrophage were combined with HiBiT Target Cells and trastuzumab to measure ADCP activity.

All HiBiT TCK Assays use one of our Bio-Glo-NB™ Luciferase detection reagents.

ADCC, TDCC and CAR-T applications:
Bio-Glo-NB™ TCK Luciferase Assay System

ADCP applications:
Bio-Glo-NB™ Lytic Luciferase Assay System