Anti-RANKL (Denosumab biosimilar - IgG1 isotype)

Human IgG1 monoclonal antibody (mAb) against human RANKL (TNFSF11)

Anti-hRANKL-hIgG1 mAb features a human IgG1 constant region (Fc) and the variable region of denosumab which targets the transmembrane and soluble forms of human RANKL (receptor activator of nuclear factor-κB ligand) antigen. RANKL is also known as member 11 of the tumor necrosis factor ligand superfamily (TNFSF11) [1]. RANKL binding to its receptor RANK induces signaling in osteoclasts and is responsible for bone resorption. RANKL-RANK also plays a pivotal role in dendritic cell maturation and T-cell differentiation [2]. 

Denosumab is a fully human IgG2 mAb that blocks the interaction of hRANKL with its receptor RANK and thus the downstream signaling.
Denosumab has been approved by the FDA for the treatment of osteoporosis and bone metastasis.

Anti-hRANKL-hIgG1 can be used in comparison with other anti-hRANKL mAbs to assess differences in the effector functions between isotypes. Human IgG1 is the most abundant immunoglobin present in serum and binds with high affinity to the Fc receptor on phagocytic cells. The human IgG1 isotype displays high complement-dependent cytotoxicity (CDC),  high antibody-dependent cellular cytotoxicity (ADCC), and very high antibody-dependent cellular phagocytosis (ADCP).

InvivoGen's Anti-hRANKL-hIgG1 is generated by recombinant DNA technology, produced in Chinese hamster ovary (CHO) cells, and purified by affinity chromatography (protein G).

More isotypes of this antibody are available and can be used for comparison of biological activities such as ADCC (see below or in the 'upon request' section).  

 Read our review on Antibody Isotypes

References:

1. Ahern E. et al., 2018. Roles of the RANKL-RANK axis in antitumour immunity -implications for therapy. Nat Rev Clin Oncol. 15(11):676-693.
2. Cheng ML. & Fong L., 2014. Effects of RANKL-Targeted Therapy in Immunity and Cancer. Front Oncol. 3:329.

Figures

Binding of Anti-hRANKL-hIgG1 mAb to coated human RANKL protein. Human RANKL (1 µg/ml) was coated on ELISA plates overnight. A 3-fold serial dilution of the Anti-hRANKL-hIgG1 mAb (red curve) or the Anti-β-Gal-hIgG1 control antibody (grey curve) was performed for the capture step. An HRP-labelled anti-hIgG antibody (1/1000 dilution) and the HRP substrate OPD (o-phenylenediamine dihydrochloride) were used for the detection step. Absorbance was read at 490 nm.

Specifications

Target: Human receptor activator of nuclear factor-κB ligand (RANKL, TNFSF11, CD254)

Clonality: Monoclonal antibody

Clone: Denosumab (anti-hRANKL-hIgG2, kappa)

Isotype: Human IgG1, kappa

Control: Human IgG1

Source: CHO cells

Formulation: 0.2 µm filtered solution in 68 mM sodium phosphate buffer (pH 7.4) with 91 mM glycine, 5% w/v saccharose, and stabilizing agents

Purity: Purified by affinity chromatography with protein G

Tested applications:  ELISA, neutralization assay

Quality control:

• Binding of Anti-hRANKL-hIgG1 to human RANKL has been confirmed using ELISA and neutralizing cellular assays.
• The complete sequence of this 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 cells.

Contents

Anti-hRANKL-hIgG1 purified monoclonal antibody (mAb) is provided azide-free and lyophilized. It is available in two quantities:

• hrankl-mab1: 100 µg
• hrankl-mab1-03: 3 x 100 µg

Anti-hRANKL-hIgG1 is shipped at room temperature.

 Upon receipt, store at -20 °C.