Human CD40L Antibody - Mouse IgG2a

Recombinant anti-human CD40L neutralizing antibody

Binding of Anti-hCD40L mAb

Anti-hCD40L-mIgG2a is a monoclonal antibody (mAb) targeting the human CD40 ligand (CD40L). This mAb specifically blocks the binding between CD40L to its receptor CD40, validated using proprietary CD40L reporter cells.  

Anti-hCD40L-mIgG2a was previously extracted from hybridoma cells (clone 13H5). It is now expressed and produced in Chinese hamster ovary (CHO) cells, ensuring reliability and lot-to-lot reproducibility. This antibody can be used together with HEK-Blue™ CD40L cells for screening and neutralization assays to block CD40L signaling induced by recombinant human CD40L (see figure).

Key features

• Each lot is functionally tested and validated.
• The complete sequence of the antibody construct has been verified.
• The absence of endotoxins is determined by the EndotoxDetect™ assay.

CD40 Ligand (CD40L) is a protein that is primarily expressed on activated T cells and is a member of the tumor necrosis factor superfamily. Of note, CD40L, together with its receptor CD40, plays a pivotal role in cellular and humoral immunity.

 More details

The hybridoma-derived Anti-hCD40L-IgG (mabg-h40l-3) antibody has been replaced by Anti-hCD40L-mIgG2a (mab10-h40l-03), which is produced by recombinant technology and purified from CHO cells.

Figures

Dose-dependent inhibition of HEK-Blue™ CD40L cell response using Anti-hCD40L-mIgG2a. Increasing concentrations of Anti-hCD40L-mIgG2a (0.1 ng/ml - 3 µg/ml) were incubated with CHO-derived recombinant human CD40L (30 ng/ml) 1 hour prior to the addition of HEK-Blue™ CD40L cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTIBlue™ Solution. Data are shown as the percentage of activity (mean ± SEM).

Specifications

Application: Neutralization assay

Isotype: Mouse IgG2a, kappa

Recommended isotype control: Mouse IgG2a

Target: CD40L

Species reactivity: Human

Clone: 13H5

Source: CHO cells

Production: Animal-free

Purification: Affinity chromatography

Physical form: Lyophilized

Formulation buffer: Sodium phosphate buffer with glycine, saccharose, and stabilizing agents

Preservative: Azide-free

Reconstitution buffer: Sterile water (not provided)

Purity: ≥ 95 %

Quality control: Each lot is functionally tested and validated.

Contents

Anti-hCD40L-mIgG2a purified monoclonal antibody is provided azide-free and lyophilized:

• mab10-h40l-03: 3 x 100 µg

 Upon receipt, store lyophilized antibody at -20 °C.

 The lyophilized product is stable for at least 1 year.

 Avoid repeated freeze-thaw cycles.

Details

CD40 Ligand (CD40L), also known as CD154, TRAP, or gp39, is a type II transmembrane glycoprotein belonging to the tumor necrosis factor (TNF) family. It is mainly expressed in CD4+-T cells and interacts with CD40 on antigen-presenting cells to regulate both humoral and cellular immune responses [1-3].

The CD40 cytoplasmic domain binds directly to several TNF receptor-associated factors (TRAFs), and this interaction is thought to initiate CD40 signaling. CD40-mediated signaling results in NF-κB, c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) activation. CD40L-CD40 interactions are thought to play an important role in the pathogenesis of many diseases [4, 5].

1. Karnell J.L. et al., 2019. Targeting the CD40-CD40L pathway in autoimmune diseases: Humoral immunity and beyond. Adv Drug Deliv Rev. 141:92-103.
2. Laman J.D. et al., 2017. Functions of CD40 and Its Ligand, gp39 (CD40L). Crit Rev Immunol. 37(2-6):371-420.
3. Elgueta R. et al., 2009. Molecular mechanism and function of CD40/CD40L engagement in the immune system. Immunol. Rev. 229, 152–172.
4. Seijkens T. et al., 2013. CD40–CD40L: Linking pancreatic, adipose tissue and vascular inflammation in type 2 diabetes and its complications. Diabetes and Vascular Disease Research. 10: 115 - 122.
5. Daoussis D. et al., 2004. Targeting CD40L: a promising therapeutic approach. Clin Diagn Lab Immunol. 11(4):635-41.