Promega - Model 4-1BB - Immune Checkpoint Bioassays Cells
From Cell Biology - Bioassays - Immune Checkpoint Bioassays
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.
Products Details
A Simpler, More Consistent 4-1BB Bioassay
- Prequalified according to ICH guidelines
- Mechanism-of-action-based assay
- No primary cell culture required
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 4-1BB Effector Cells included are provided in a thaw-and-use format as cryopreserved cells that can be thawed, plated and used in an assay without the need for cell propagation.
The 4-1BB Effector Cells have been engineered to express the response element (RE) upstream of luc2P, as well as exogenous 4-1BB receptor. When 4-1BB binds to 4-1BB Effector Cells, the receptor transduces intracellular signals resulting in luminescence.The bioluminescent signal is detected and quantified using Bio-Glo™ Luciferase Assay System (Cat.# G7940) and a standard luminometer such as the GloMax® Discover System.
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.
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