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Non-fucosylated Anti-hTIGIT mAb

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Anti-hTIGIT-hIgG1fut

Recombinant monoclonal human IgG1fut antibody against human TIGIT

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100 µg

htigit-mab13

Non-fucosylated recombinant Anti-hTIGIT mAb

 TIGIT in the context of immunotherapy
TIGIT in the context of immunotherapy

Anti-hTIGIT-hIgG1fut is a recombinant monoclonal antibody (mAb) featuring a variable region that recognizes human TIGIT and a non-fucosylated constant region of the human IgG1 (hIgG1fut) isotype.

TIGIT (T cell immunoglobulin and ITIM domain) is an inhibitory checkpoint that has been implicated in tumor immunosurveillance [1]. 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 [4].

Features of Anti-hTIGIT-hIgG1fut:

  • Targets specifically human TIGIT
  • Features the hIgG1fut constant region for enhanced effector function
  • Functionally validated by flow cytometry

Anti-hTIGIT-hIgG1fut is a non-fucosylated antibody, whereby the absence of fucose residues from N-glycans of the IgG-Fc results in an enhancement of antibody-dependent cellular cytotoxicity (ADCC). This increased ADCC is without any detectable change to complement-dependent cytotoxicity (CDC) or antigen-binding capability. Anti-hTIGIT-hIgG1fut was generated by recombinant DNA technology, produced in CHO cells (deficient for fucosylation), and purified by affinity chromatography with protein G.

 

References:

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.

Figures

Binding of Anti-hTIGIT-hIgG1fut to target cells
Binding of Anti-hTIGIT-hIgG1fut to target cells

Anti-hTIGIT-hIgG1fut (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
Comparison of ADCC potency for native and engineered anti-human TIGIT antibody isotypes

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.

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Specifications

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 (deficient for fucosylation)

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-hIgG1fut binds and blocks ligand activation of TIGIT found on the surface of cells

Quality control:

  • Binding of Anti-hTIGIT-hIgG1fut 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.
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Contents

  • 100 µg Anti-hTIGIT-hIgG1fut purified antibody (provided azide-free and lyophilized)

room temperature Product is shipped at room temperature.

store Store lyophilized antibody at -20 °C.

stability Lyophilized product is stable for at least 1 year

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This antibody has been removed from our catalog. However, you may contact us if you wish to order minimal quantities.

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