Non-glycosylated Anti-hTIGIT mAb
|Anti-hTIGIT-hIgG1NQ||Unit size||Cat. code||Docs||Qty||Price|
Recombinant monoclonal human IgG1NQ antibody against human TIGIT
Non-glycosylated recombinant Anti-hTIGIT mAb
Anti-hTIGIT-hIgG1NQ is a recombinant monoclonal antibody (mAb) featuring a variable region that recognizes human TIGIT and a non-glycosylated constant region of the human IgG1 (hIgG1NQ) 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-hIgG1NQ:
- Targets specifically human TIGIT
- Features the hIgG1NQ constant region for abrogated effector function
- Functionally validated by flow cytometry
Anti-hTIGIT-hIgG1NQ contains an N-glycosylation mutation in the constant region of human IgG1. Potential asparagine (N) glycosylation sites are substituted by glutamine (Q) residues, resulting in the production of a non-glycosylated antibody. Glycosylation of an antibody has no effect on antigen binding but is essential for Fc receptor-mediated activity, and therefore, the effector function of Anti‑hTIGIT‑hIgG1NQ is severely compromised. Anti-hTIGIT-hIgG1NQ 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-hIgG1NQ (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.
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-hIgG1NQ binds and blocks ligand activation of TIGIT found on the surface of cells
- Binding of Anti-hTIGIT-hIgG1NQ 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-hIgG1NQ 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