Anti-PD-1 (Nivolumab biosimilar - IgG1 isotype)

Principle of PD-1/PD-L1 cellular assay
(click to enlarge)

Human IgG1 monoclonal antibody (mAb) against human PD-1

Anti-hPD1-Ni-hIgG1 features the constant region of the human IgG1 isotype and the variable region of nivolumab. Nivolumab is a fully human IgG4 (S228P) monoclonal antibody that targets the programmed cell death 1 (PD-1) receptor found on activated T cells, B cells, and myeloid cells. Under normal physiological conditions, PD-1 negatively regulates T cell activation thereby preventing autoimmunity [1]. Under pathological conditions, cancer cells produce PD-L1 (programmed cell death 1 ligand 1), the agonist that binds and activates PD-1. Activated PD-1 enables the cancer cells to evade the immune system. Nivolumab binds and blocks the activation of the PD-1 receptor, thereby resulting in the activation of T cells and cell-mediated immune responses [2, 3]. This antibody contains an engineered hinge region mutation (S228P) designed to prevent exchange of IgG4 molecules. Nivolumab has been approved by the FDA for the treatment of melanoma and metastatic squamous non-small cell lung cancer (NSCLC).
Anti-hPD1-Ni-hIgG1 was generated by recombinant DNA technology. It has been produced in Chinese hamster ovary (CHO) cells and purified by affinity chromatography with protein G.

References:

McDermott D. & Atkins M., 2013. PD-1 as a potential target in cancer therapy. Cancer Med. 2: 662–73.
Wang C. et al., 2014. In vitro characterization of the anti-PD-1 antibody nivolumab, BMS- 936558, and in vivo toxicology in non-human primates..Cancer Immunol Res. 2:846-56.
Gunturi A. & McDermott DF., 2015. Nivolumab for the treatment of cancer. Expert Opin Investig Drugs. 24:253-60.

Figures

Comparison of ADCC potency for native and engineered anti-human PD-1 antibody isotypes: Raji-hPD-1 cells were incubated with gradient concentrations of Anti-hPD-1 or Anti-β-galactosidase (β-gal) mAbs for 1 hour. Jurkat-Lucia™ NFAT-CD16 effector cells were then co-incubated with targets 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.

Activation of Jurkat-Lucia™ TCR-hPD-1 cells.
Raji-APC-hPD-L1 and Jurkat-Lucia™ TCR-hPD-1 cells were incubated with gradient concentrations of AntihPD-1 hIgG1 (Ni: Nivolumab variable region; Pem: Pembrolizumab variable region) or Anti-hPD-L1 mAbs for 6 hours. NFAT activation, reflecting the disruption of PD-1/PD-L1 inhibitory interaction, was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™.
The fold increase over non induced cells (no mAbs) is shown.

Specifications

Specificity: Targets cells expressing human PD-1

Clonality: Monoclonal antibody

Clone: Nivolumab (Anti-hPD1-hIgG4 (S228P), kappa)

Isotype: Human IgG1, kappa

Control: Human IgG1, kappa

Source: CHO cells

Purity: Purified by affinity chromatography with protein G

Applications: Anti-hPD1-Ni-hIgG1 binds and blocks ligand activation of the PD-1 receptor on the surface of activated T cells.

Quality control:

• Binding of Anti-hPD1-Ni-hIgG1 to human PD-1 has been tested using flow cytometry.
• 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-hPD1-Ni-hIgG1 purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• hpd1ni-mab1: 100 µg
• hpd1ni-mab1-03: 3 x 100 µg

Product is shipped at room temperature.

 Upon receipt, store at -20°C.