|Anti-hTIGIT-hIgG1||Unit size||Cat. code||Docs||Qty||Price|
Recombinant monoclonal human IgG1 antibody against human TIGIT
Recombinant Anti-hTIGIT mAb
Anti-hTIGIT-hIgG1 is a recombinant monoclonal antibody (mAb) featuring a variable region that recognizes human TIGIT, and the constant region of the human IgG1 (hIgG1) isotype.
TIGIT (T cell immunoglobulin and ITIM domain) is an inhibitory checkpoint that has been implicated in tumor immunosurveillance . TIGIT is specifically expressed on immune cells including, natural killer (NK) cells and a range of T cell subsets. TIGIT binds to CD155 (PVR) and CD112 (PVRL2, nectin-2), which are expressed on antigen-presenting cells (APCs), T cells, and a variety of non-hematopoietic cells including tumor cells. Interestingly, TIGIT competes with CD226 (also known as DNAM-1) and CD96 (also known as Tactile) for the same ligands [1,2]. Upon binding to its ligand, phosphorylation of TIGIT inhibits the NF-κB, P13K, and MAPK pathways, and leads to a strong reduction of NK cytotoxicity as well as inhibition of T cell activation, proliferation, and effector functions [2,3].
The blockade of TIGIT is highly favorable in cancer immunotherapy due to a number of reasons including its low expression in peripheral lymphoid organs and high expression in tumor-infiltrating lymphocytes (TILs), the established synergy of TIGIT with other co-inhibitory immune checkpoints, and its ligands being widely expressed on tumor cells [1,2]. The dual blockade of TIGIT and PD-L1 has shown synergistic effects in a murine tumor model, resulting in complete tumor rejection and induced protective memory responses. A similar synergistic effect has been noted with PD-1 and Tim-3 [1,2]. Interestingly, TIGIT’s role in the tumor microenvironment (TME) may also be intertwined with the microbiome. The suppressive function of TIGIT is also exploited by a bacterium commonly found in the TME Fusobacterum nucleatun, to inhibit protective immune responses .
Features of Anti-hTIGIT-hIgG1:
- Targets specifically human TIGIT
- Features the hIgG1 constant region for high effector function
- Functionally validated by flow cytometry
Anti-hTIGIT-hIgG1 contains the human IgG1 constant region, which is known to bind with high affinity to the Fc receptor on phagocytic cells. Therefore, Anti-hTIGIT-hIgG1 displays high effector function, including antibody‑dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Anti-hTIGIT-hIgG1 was generated by recombinant DNA technology, produced in CHO cells, and purified by affinity chromatography with protein G.
1. Solomon, B. L. et al, 2018. TIGIT: a novel immunotherapy target moving from bench to bedside. Cancer Immunol Immunother 67, 1659-1667.
2. Anderson, A.C. et al. 2016. Lag-3, Tim-3, and TIGIT: Co-inhibitory Receptors with Specialized Functions in Immune Regulation. Immunity 44, 989-1004.
3. Joller, N. et al. 2011. Cutting edge: TIGIT has T cell-intrinsic inhibitory functions. J Immunol 186, 1338-1342.
4. Gur, C. et al. 2015. Binding of the Fap2 protein of Fusobacterium nucleatum to human inhibitory receptor TIGIT protects tumors from immune cell attack. Immunity 42, 344-355.
Anti-hTIGIT-hIgG1 (2 μg) was added to Raji-null (negative control) and Raji-hTIGIT cells (500 000 cells/ml) and incubated at room temperature for 30 minutes. Following this, a secondary antibody, PE, was added and incubated again at room temperature for 30 minutes. The binding affinity was then measured using flow cytometry.
Comparison of ADCC potency for native and engineered anti-human TIGIT antibody isotypes: Raji-hTIGIT cells were incubated with gradient concentrations of Anti-hTIGIT or Anti-β-galactosidase (β-gal) mAbs for 1 hour. Jurkat-Lucia™ NFAT-CD16 effector cells were then co-incubated with target cells for 6 hours. NFAT activation, reflecting the induced ADCC response, was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™. Percentages of the maximal response normalized to the IgG1 isotype are shown.
Specificity: Targets cells expressing human TIGIT (T cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain)
Clonality: Monoclonal antibody
Isotype: Human IgG1, kappa
Source: CHO cells
Formulation: 0.2 μm filtered solution in a sodium phosphate buffer with glycine, saccharose, and stabilizing agents.
Purification: Purified by affinity chromatography with protein G
Applications: Anti-hTIGIT-hIgG1 binds and blocks ligand activation of TIGIT found on the surface of cells
- Binding of Anti-hTIGIT-hIgG1 to human TIGIT on cells has been validated using flow cytometry.
- The complete sequence of the antibody has been verified.
- The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cellular assays.
- 100 µg Anti-hTIGIT-hIgG1 purified antibody (provided azide-free and lyophilized)
Product is shipped at room temperature.
Store lyophilized antibody at -20 °C.
Lyophilized product is stable for at least 1 yearBack to the top