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Anti-VISTA (Biosimilar antibody - IgG1fut isotype)

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Anti-hVISTA-hIgG1fut

Human VISTA biosimilar antibody - human IgG1, non-fucosylated

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100 µg

3 x 100 µg

hvista-mab13
+-
$109
  • About
  • Specifications
  • Contents
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Non-fucosylated human IgG1 monoclonal antibody (mAb) against human VISTA

 VISTA in the context of immunotherapy
VISTA in the context of immunotherapy

Anti-hVISTA-hIgG1fut is a recombinant monoclonal antibody (mAb) featuring a variable region that recognizes human VISTA, a member of the CD28/B7 protein superfamily, and a non-fucosylated constant region of the human IgG1 (hIgG1fut) isotype.

V-domain Ig-containing suppressor of T-cell activation (VISTA), also known as programmed death-1 homolog (PH-1H) is a known immune checkpoint, that directly suppresses T cell activation, proliferation, and cytokine production [1]. VISTA can be considered both a ligand and a receptor, whereby it transmits inhibitory signals to T cells, either extrinsically, through expression on myeloid cells (i.e. macrophage and dendritic cells) and binding to an unknown receptor on T cells, or intrinsically, through its expression on T cells (i.e. CD4+, CD8+) [2]. In the context of cancer immunotherapy, VISTA has been found to be heightened on cells that infiltrate the tumor microenvironment (TME) and not the tumor cells themselves, therefore, VISTA-blocking antibodies are of interest. These function to inhibit the engagement of VISTA with its receptor on T cells or by directly targeting VISTA expressed on naïve and effector T cells, preventing it from providing negative signaling [1, 2].  

Features of Anti-hVISTA-hIgG1fut:

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

Anti-hVISTA-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-hVISTA-hIgG1fut was generated by recombinant DNA technology, produced in CHO cells (deficient for fucosylation), and purified by affinity chromatography with protein G.

 

References:

1. Nowak, E.C. et al. 2017. Immunoregulatory functions of VISTA. Immunol Rev 276, 66-79.
2. Xu, W. et al. 2018. The structure, expression, and multifaceted role of immune-checkpoint protein VISTA as a critical regulator of anti-tumor immunity, autoimmunity, and inflammation. Cell Mol Immunol 15, 438-446.

Figures

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

Anti-hVISTA-hIgG1fut (1 μg) was added to Raji-null (negative control) and Raji-hVISTA 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.

ADCC assay with Raji-hVISTA cells
ADCC assay with Raji-hVISTA cells

Comparison of ADCC potency for native and engineered anti-human VISTA antibody isotypes: Raji-hVISTA cells were incubated with gradient concentrations of Anti-hVISTA 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 VISTA (V-domain Ig containing suppressor of T-cell activation)

Clonality: Monoclonal antibody

Isotype: Human IgG1fut, kappa

Control: Human IgG1fut 

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-hVSITA-hIgG1fut binds and blocks ligand activation of the VISTA receptor found on the surface of cells

Quality control:

  • Binding of Anti-hVISTA-hIgG1fut to human VISTA 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

Anti-hVISTA-hIgG1fut purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

  • hvista-mab13: 100 µg
  • hvista-mab13-03: 3 x 100 µg

room temperature Product is shipped at room temperature.

store Upon receipt, store at -20 °C.

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