Human PD-1 (CD279) Antibody - Nivolumab IgG1fut isotype

Anti-human PD-1 antibody - Human IgG1 non-fucosylated (high effector functions)

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

Anti-hPD1-Ni-hIgG1fut features the variable region of nivolumab and a non-fucosylated IgG1 constant region for high effector functions. Nivolumab is a therapeutic, 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 the 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-hIgG1fut is a non-fucosylated antibody. The absence of the fucose residue from the N-glycans of IgG-Fc results in dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) without any detectable change in complement-dependent cytotoxicity (CDC) or antigen-binding capability [4, 5]. It has been produced in Chinese hamster ovary (CHO) cells that are deficient for fucosylation and purified by affinity chromatography with protein G.

References:

1. McDermott D. & Atkins M., 2013. PD-1 as a potential target in cancer therapy. Cancer Med. 2: 662–73.
2. 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.
3. Gunturi A. & McDermott DF., 2015. Nivolumab for the treatment of cancer. Expert Opin Investig Drugs. 24:253-60.
4. Yamane-Ohnuki N. & Satoh M., 2009. Production of therapeutic antibodies with controlled fucosylation.corresponding MAbs. 1(3): 230–236.
5. Mizushima T., 2011. Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. Genes Cells. 16(11): 1071–1080

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.

Specifications

Application: Cellular assay, ELISA, flow cytometry, Fc interaction studies

Isotype:  Human IgG1fut (non-fucosylated), kappa

Recommended isotype control: Human IgG1fut

Target: Human PD-1

Species reactivity: Human

Clone: Nivolumab, BMS-936558, ONO-4538, MDX-1106

Sterility: 0.2 µm filtration

Source: CHO cells 

Production: Animal-free

Purification: Protein A

Molecular weight: 

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

• 3 x 100 µg Anti-hPD1-Ni-hIgG1fut provided azide-free and lyophilized

Product is shipped at room temperature.

 Upon receipt, store at -20°C.