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Murine Anti-mCTLA4 mAb

Anti-mCTLA4-mIgG2a InvivoFit™ Unit size Cat. code Docs Qty Price
9D9-derived mouse monoclonal antibody against murine CTLA-4
1 mg
10 mg
mctla4-mab10-1
+-
$346.00

Recombinant mouse mAb against murine CTLA-4 for in vivo use

Effect of Anti-CTLA4 on T cell function
Effect of Anti-CTLA4 on T cell activation

Anti-mCTLA4-mIgG2a InvivoFit™ is a mouse anti-mouse monoclonal antibody (mAb) featuring the variable region of the previously described anti-mCTLA-4 9D9 mAb [1] and a murine IgG2a constant region. The 9D9 mAb has a sequence that is 100% murine (constant and variable regions), ensuring minimal immunogenicity upon repeated injections into mice. This mAb targets murine cytotoxic T-lymphocyte-associated protein 4 (mCTLA-4) and blocks CTLA4-mediated negative signals downregulating T cell activation.  Thus, anti-mCTLA-4 mAb unleashes T cell activity [1, 2].
Notably, the 9D9 mAb, along with most commercially available anti-mCTLA4 mAbs feature the IgG2b isotype which displays low antitumor activity in vivo [2]. To overcome this issue, Anti-mCTLA4-mIgG2a InvivoFit™ was generated by recombinant technology and features the murine IgG2a isotype, which mediates potent cytotoxic functions. Anti-mCTLA4-mIgG2a is provided in an InvivoFit grade, a high-quality standard specifically adapted to in vivo studies.

 

Key features of Anti-mCTLA4-mIgG2a InvivoFit™:

  • Sequence is 100% murine
  • Features the IgG2a isotype (associated with high antitumor activity)
  • Guaranteed sterile, endotoxin level < 1 EU/mg
  • Specifically designed for in vivo studies in mice
  • Low aggregation < 5%
  • Produced in animal-free facilities and defined media


Anti-mCTLA4-mIgG2a InvivoFit™ is produced in Chinese hamster ovary (CHO) cells, purified by affinity chromatography with protein A, and it's binding to mCTLA-4 is validated using ELISA.

 

References:

1. Selby MJ. et al., 2013. Anti-CTLA-4 antibodies of IgG2a isotype enhance antitumor activity through reduction of intratumoral regulatory T cells. Cancer Immunol Res. 1(1):32-42.
2. Ribas A. & Wolchock J.D., 2018. Cancer immunotherapy using checkpoint blockade. Science. 359:1350-55.

Figures

Evaluation of Anti-mCTLA4-mIgG2a binding to mCTLA-4
Evaluation of Anti-mCTLA4-mIgG2a binding to mCTLA-4

ELISA binding of Anti-mCTLA4-mIgG2a InvivoFit™ to recombinant murine CTLA-4.

A dilution series of Anti-mCTLA4-mIgG2a InvivoFit™ and anti-mCTLA4 (9D9 clone) was used for the detection of coated mCTLA-4 antigen at 5  µg/ml. Detection of bound antibody was performed using an HRP‑conjugated anti‑murine κ-light chain secondary antibody.  HRP activity was assessed by a colorimetric assay using Tetramethylbenzidine (TMB) substrate. Optical density was measured at 490 nm. Data are shown as percentage absorbance.

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Specifications

Specificity: Targets cells expressing murine CTLA-4

Formulation: Lyophilized from 0.2 μm filtered solution in 150 mM sodium chloride, 20 mM sodium phosphate buffer with 5% saccharose

Clonality: Monoclonal antibody

Isotype: Murine IgG2a, kappa

Source: CHO cells

Purity: Purified by affinity chromatography with protein A

Tested application: ELISA

Quality control:

  • Binding confirmed using ELISA
  • The complete sequence of this antibody has been verified
  • < 5% aggregates (confirmed by size exclusion chromatography)
  • Guaranteed sterile and <1 EU/mg (determined by the LAL assay)
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Contents

Anti-mCTLA4-mIgG2a InvivoFit™ is provided sterile, endotoxin-free, azide-free and lyophilized.  

This product is available in two pack sizes:

  • mctla4-mab10-1​: 1 mg
  • mctla4-mab10-10​: 10 mg

 

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

Alert Avoid repeated freeze-thaw cycles.

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Details

The cytotoxic T-lymphocyte-associated protein 4 (CTLA-4, also known as CD152) is an inhibitory receptor expressed on activated T lymphocytes. T-cell activation requires two signals. Signal 1 is delivered upon interaction of the T cell receptor (TCR) with antigenic peptides bound to major histocompatibility complex (MHC) molecules on antigen-presenting cells (APCs). Signal 2 is delivered upon interaction of the co-stimulatory receptor CD28 with the B7 family ligands, B7-1 (CD80) and B7-2 (CD86), on APCs.
CTLA-4 is an immune checkpoint molecule upregulated upon T cell activation. It exerts competitive binding to CD28 ligands, thereby preventing T-cell overstimulation. CTLA-4 blockade with monoclonal antibodies (mAbs) has been used to treat patients with metastatic melanoma.

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In vivo data

In vivo efficacy of InvivoGen’s Anti-mCTLA4-mIgG2a
In vivo efficacy of InvivoGen’s Anti-mCTLA4-mIgG2a

In vivo efficacy of InvivoGen’s Anti-mCTLA4-mIgG2a

Unlike most immune checkpoint (IC) mAbs on the market, which are produced in hybridomas, InvivoGen’s mouse anti-mouse mAbs are expressed in virus-free CHO cells. This ensures structural stability as well as guaranteeing the highest purity and batch-to-batch consistency; important for mAb efficacy in vivo (Figure 1). 

BALB/c mice were challenged with the murine colorectal carcinoma cell line, CT26. After 8 days, treatment was started with InvivoGen's Anti‑mCTLA4‑mIgG2a (recombinant mAb), Competitor X’s Anti‑mCTLA4 (produced in hybridomas; clone 9D9-mIgG2b), or negative controls (saline only and isotype control mAb). Injections were performed twice a week for 3 weeks (a total of 6 times). After 60 days of monitoring, (A) 7/10 mice were tumor-free in the Competitor X mAb-treated group, compared to (B) 9/9 mice tumor-free in the InvivoGen mAb-treated group. Additionally, no significant differences were observed in the health of the mice, with comparable body weight throughout the study (C). Most notably, after 6 injections, 30%* of the mice in the Competitor X mAb-treated group were euthanized due to excessive tumor volume (endpoint criteria), whereas complete tumor regression was observed in all mice in the InvivoGen mAb-treated group (D)

In summary, InvivoGen’s optimized Anti‑mCTLA4‑mIgG2a shows increased in vivo efficiency compared to Competitor X’s mAb, which features the commonly used IgG2b isotype, known to display low antitumor activity in vivo. 

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