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Promega - Model 4-1BB - Immune Checkpoint Bioassays Cells
4-1BB (CD137/TNFRSF9), a member of the tumor necrosis factor receptor superfamily, is an inducible co-stimulatory receptor expressed on T cells, natural killer (NK) cells and innate immune cell populations. When present on the cell surface, 4-1BB interacts with 4-1BB ligand (4-1BBL) and induces subsequent cell proliferation and production of interferon gamma (IFNγ) and interleukin-2 (IL-2), particularly in T and NK cells.
A Simpler, More Consistent 4-1BB Bioassay
- Prequalified according to ICH guidelines
- Mechanism-of-action (MOA)-based assay
- No primary cell culture required
- Supplied in convenient thaw-and-use format; also available in cell propagation model (CPM) format
Current methods used to measure the activity of biologic drugs targeting 4-1BB rely on assays that are laborious and highly variable due to their reliance on donor primary cells, complex assay protocols and unqualified assay reagents and, as a result, are difficult to establish in a quality-controlled, drug-development setting.
The 4-1BB Bioassay is a bioluminescent cell-based assay that overcomes the limitations of existing assays and can be used to measure the potency and stability of ligands or agonist antibodies that can bind and activate 4-1BB. The assay consists of a genetically engineered Jurkat T cell line that expresses human 4-1BB and a luciferase reporter driven by a response element that can respond to 4-1BB ligand/agonist antibody stimulation. The 4-1BB Effector Cells are provided in thaw-and-use format as cryopreserved cells that can be thawed, plated and used in an assay without the need for cell culture and propagation.
The assay is also available in CPM format (Cat.# J2332) containing cryopreserved cells that can be thawed, propagated and banked for long-term use.
Standardized Reagents Improve Results
- Thaw-and-use cell format provides significant time and labor savings over traditional methods while reducing variability.
- Kit includes all of the required reagents in standardized formats.
- Easily amenable to high-throughput workflows.
Panel A. Assay with FcγRIIb-dependent agonist antibody. The bioassay consists of two engineered cell lines, 4-1BB Effector Cells and FcγRIIb CHO-K1 Cells. In the presence of FcγRIIb CHO-K1 Cells, the anti-4-1BB antibody can be crosslinked, thereby inducing 4-1BB pathway-activated luminescence. Panel B. Assay with FcγRIIb-independent agonist antibody or ligand. The bioassay consists of one engineered cell line, 4-1BB Effector Cells. In the absence of agonist antibody or 4-1BB ligand, the 4-1BB receptor is not activated and luminescence signal is low. The addition of agonist antibody or 4-1BB ligand induces the 4-1BB pathway-activated luminescence, which can be detected in a dose-dependent manner.
Accuracy and Precision are critical to bioassays and the development of biologics. The 4-1BB Bioassay is both accurate and precise. Accuracy is a measure of how close results are to a target or reference value. Precision is a measure of repeatability. This assay was developed and prequalified according to ICH guidelines for accuracy and precision giving it the performance characteristics required for many applications of a bioassay used for antibody drug discovery research and development.