T Cell Activation Bioassay (NFAT) 5X
Immunotherapy strategies aimed at inducing, strengthening or engineering T cell responses have emerged as promising approaches for the treatment of diseases such as cancer and autoimmunity. T cell activation is initiated by engagement of the TCR/CD3 complex and the co-stimulatory receptor CD28, with TCR/CD3 engagement activating the NFAT pathway and TCR/CD3 + CD28 co-engagement activating NFAT, AP-1 and NF-kappaB pathways and inducing IL-2 production. The T Cell Activation Bioassays are bioluminescent cell-based assays that overcome the limitations of existing assays and can be used for the discovery and development of novel biologics such as bispecific antibodies and CAR-T cell therapies. The assays consist of a genetically engineered Jurkat T cell line that expresses a luciferase reporter (TCR/CD3 Effector Cells) driven by either an NFAT-response element (NFAT-RE) or an IL-2 promoter. When the TCR/CD3 Effector Cells (NFAT) are engaged with an appropriate TCR/CD3 ligand or anti-TCR/CD3 antibody, the TCR transduces intracellular signals resulting in NFAT-RE-mediated luminescence. Similarly, when the TCR/CD3 Effector Cells (IL-2) are co-engaged with an anti-TCR/CD3 and an anti-CD28 stimulus, receptor-mediated signaling results in IL-2 promoter-mediated luminescence. The bioassay is prequalified according to ICH guidelines and shows the precision, accuracy and linearity required for routine use in potency and stability studies. The bioassay workflow is simple, robust and compatible with 96-well and 384-well plate formats used for antibody screening and drug discovery. Additionally, the bioassay is tolerant to human serum, indicating potential for further development into a neutralizing antibody bioassay. Product Kit Formats: The T Cell Activation Bioassay Kits include TCR/CD3 Effector Cells (NFAT or IL-2), Cell Culture Medium and Assay Reagents. The kits are available in 1X and 5X sizes.