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Applications: Screen or titrate small molecule inhibitors or antibodies that block CD155 binding to TIGIT for drug discovery and high-throughput screening (HTS) applications.
Background: TIGIT (T-cell immunoreceptor with Ig and ITIM domains) is an immune checkpoint, co-inhibitory receptor of the family of PVR (poliovirus receptor)-like proteins that is highly expressed on the surface of Natural Killer (NK) cells, activated CD4+, CD8+, and regulatory T cells. TIGIT has three ligands: CD155, CD112 and CD113. TIGIT has the highest affinity for CD155, which is present on antigen presenting cells (APCs) such as dendritic cells, on T cells, B cells, and macrophages. TIGIT binding to CD155 recruits the Src homology (SH) domain-containing protein tyrosine phosphatases SHP1 and SHP2 or the inositol phosphatases SHIP1 and SHIP2 to the TIGIT ITIM domain (immunoreceptor tyrosine-based inhibitory motif). This increases the production of IL-10 and decreases IL-12 release, and it suppresses NF-kB (nuclear factor kappa-light chain enhancer of activated B cells) and NFAT (nuclear factor of activated T cells) T cell receptor (TCR) signaling, which blocks T cell proliferation and cytokine production. TIGIT may also play a role in immune regulation via cell-intrinsic mechanisms, with anti-TIGIT agonist antibodies blocking the anti-CD3/anti-CD28 activation of T cells and DNAM-1 (DNAX accessory molecule-1) signaling in the absence of APCs. TIGIT is found at high levels in TILs (tumor-infiltrating lymphocytes) in AML (acute myeloid leukemia), MM (multiple myeloma), NSCLC (non-small cell lung carcinoma) and other cancers. On the other hand, cancer cells can also express CD155. The development of TIGIT-blocking strategies is an active area of research, with several ongoing clinical trials using anti-TIGIT blocking antibodies.
Contraindications: The DMSO concentration in the final reaction should be ‰¤1%.
Description: The CD155:TIGIT [Biotinylated] Inhibitor Screening Chemiluminescence Assay Kit is an ELISA-based assay designed for screening and profiling molecules that block the binding between CD155 (cluster of differentiation 55) and TIGIT (T-cell immunoreceptor with Ig and ITIM domains). This kit comes in a convenient 96-well format, with enough recombinant human biotin-labeled TIGIT (amino acids 22-141), CD155 (amino acids 27-343), streptavidin-labeled HRP, and assay buffer for 100 reactions.A 96-well plate is coated with the human recombinant CD155 protein. After blocking, the plate is pre-incubated with an inhibitor or neutralizing antibody. After incubation with Biotin-TIGIT, the plate is washed and Streptavidin-HRP is added. The ELISA ECL substrate is added, and the resulting signal is measured using a chemiluminescence microplate reader. The chemiluminescence signal is proportional to the binding of CD155 to TIGIT.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Uniprot: TIGIT: Q495A1; CD155: P15151
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
References: Yu X., et al., 2009. Nat. Immunol. 10(1): 48-57.
Stanietsky N., et al., 2009. Proc. Natl. Acad. Sci. 106(42): 17858-17863.
Harjunpaa H. and Guillerey C., 2020 Clin Exp Immunol 200(20:108-119.
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