PBMC ADCC Bioassay

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MOA-Based Functional Bioassay to Measure Fc Effector Activity

  • Human PBMCs, ADCC-qualified
  • Simple, add-mix-read format
  • Choice of target cells expressing HiBiT fusion protein and corresponding PBMC ADCC Bioassay Kit (see "Available Assay Formats" below)

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PBMC ADCC Bioassay

Eliminate the Uncertainty and Variability of Primary Cell-Based ADCC Assays

The peripheral blood mononuclear cell (PBMC)-based assay provides improved consistency and robustness for measuring antibody-dependent cellular cytotoxicity (ADCC). The assays provide ADCC-qualified PBMC effector cells and a choice of popular target cells expressing a HiBiT fusion protein. Upon target-cell killing, a bright luminescent signal is generated. The bioassay is simple, homogenous, highly sensitive and provides a robust assay window. It demonstrates antibody potency comparable with the ADCC Reporter Bioassay in ADCC bridging studies.

Available Assay Formats

Effector Cells Target Cells
(HiBiT Fusion)
Bioassay Kits*
Human PBMCs, ADCC-Qualified Raji (HaloTag®-HiBiT) PBMC ADCC Bioassay Kit (Raji)
Ramos (HaloTag®-HiBiT) PBMC ADCC Bioassay Kit (Ramos)
A549 (HaloTag®-HiBiT) PBMC ADCC Bioassay Kit (A549)
H929 (HaloTag®-HiBiT) PBMC ADCC Bioassay Kit (H929)
SK-BR-3 (HaloTag®-HiBiT) PBMC ADCC Bioassay Kit (SK-BR-3)
*Each Bioassay Kit includes Human PBMCs, ADCC-Qualified, the selected Target Cells, RPMI 1640 Media, Fetal Bovine Serum and the Nano-Glo® HiBiT Extracellular Detection System.

Target Cell Information

Target Cells Cell Line Background Drug Targets Potential Applications Test Drugs Available*
Raji B lymphoma CD19, CD20, CD22 mAb, ADC, Bispecific, CAR-T rituximab, daratumumab (ADCC)
blinatumomab (TDCC)
Ramos B lymphoma CD19, CD20, CD22 mAb, ADC, Bispecific, CAR-T rituximab, daratumumab (ADCC)
blinatumomab (TDCC)
H929 Multiple myeloma BCMA ADC, Bispecific, CAR-T daratumumab (ADCC)
A549 Lung carcinoma EGFR mAb, Bispecific, CAR-T cetuximab, trastuzumab, daratumumab (ADCC)
SK-BR-3 Breast cancer HER2, EpCAM  mAb, Bispecific, CAR-T trastuzumab (ADCC)

*Drugs are commercially available for the following targets: Blinatumomab: CD3xCD19 BiTE; Rituximab: anti-CD20; Daratumumab: anti-CD38; Cetuximab: anti-EGFR; Trastuzumab: anti-HER2.

Assay Principle

When HiBiT-expressing target cells are incubated with an antibody and PBMCs, the target cells are lysed and release HiBiT proteins. These proteins bind to the complementary polypeptide LgBiT in the detection reagent to reconstitute functional NanoBiT® luciferase and generate luminescence.

Principle of the PBMC ADCC bioassay

Comparison Between PBMC and ADCC Reporter Bioassay

The PBMC ADCC Bioassay, using engineered HiBiT target cells (left panel), measures the antibody potency and specific target cell killing for anti-CD20 antibody rituximab. The results are comparable to those obtained with the Human ADCC Reporter Bioassay, V Variant (right panel).

PBMC ADCC assay Raji cells
ADCC Reporter Bioassay Raji target cells

Product Format

  • Includes Human PBMC, ADCC-Qualified, HaloTag®-HiBiT Target Cells, FBS, Cell Culture Medium and Nano-Glo® HiBiT Extracellular Detection System.
  • Recommended for use as a starter kit.
  • PBMCs, ADCC-Qualified available in frozen format.
  • Target Cells (Raji, Ramos, A549, H929 and SK-BR-3) available in propagation model or thaw and use formats.
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