Anti-PD-1 [5C4.B8 (Nivolumab)]

Absolute Antibody

Anti-PD-1 [5C4.B8 (Nivolumab)]

7

Nivolumab binds to the extracellular portion of human PD-1 (2.6 nM - Scatchard analysis and SPR) - the antibody also binds to cynomolgus PD-1 with a similar affinity (3.9 nM - SPR). The antibody does not bind to other immunoglobulin superfamily proteins s

Nivolumab has been shown to bind to PD-1-expressing CHO cells (EC50 ~1.66nM). Nivolumab binds CD4+ T cells (EC50 ~0.64 nM) and stains only memory and effector, and not naiive CD4+ or CD8+ T cells from human peripheral blood by FC. The antibody is able to block the interaction between PD-1 and its ligands PDL-1 and PDL-2 (IC50 ~2.52 nM and ~2.59 nM, respectively - determined by SPR) - these IC50 values are also similar to that measured by FACS to evaluate ligand binding to PD-1 expressed on CHO cells. In an allogenic T-cell/DC MLR, Nivolumab-mediated inhibition of PD-1 results in enhancement of IFNgamma release, and also enhances IL-2 secretion (97-139% over an isotype control) in response to the superantigen SEB using human peripheral blood mononuclear cells. The same is also observed in a CMV-restimulation assay. Nivolumab at very low concentrations (~1.5 ng/mL) is able to enhance T-cell reactivity in the presence of a T-cell receptor stimulus - nivolumab has no stimulatory effect in the absence of antigen or T-cell receptor stimulus. In the therapeutically used human IgG4 (S228P) format, this antibody is unable to mediate ADCC (antibody-dependent cell-mediated cytotoxicity) or CDC (complement-dependent cytotoxicity).

Flow Cytometry, ImmunoHistoChemistry, Neutralisation/Blocking, Other Application

IHC; FC; SPR; block

Research Use Only

Monoclonal

5C4.B8 (Nivolumab)

PDCD1

programmed cell death 1

CD279; hPD-1; hPD-l; hSLE1; PD1; PD-1; programmed cell death 1 protein; programmed cell death protein 1; protein PD-1; SLEB2; systemic lupus erythematosus susceptibility 2

Human

IgG4

Programmed cell death protein 1

Human, Monkey

Human, cynomolgus

mAb PD1.5 was prepared by immunizing IgH and IgK knock-out transgenic mice possessing a human immunoglobulin (heavy chain) minilocus with recombinant human PD-1-Fc protein consisting of the extracellular domain of PD-1 (amino acids 1–167) and the Fc portion of human IgG1, and Chinese hamster ovary (CHO) cells expressing human PD-1. Nivolumab was generated by grafting the variable regions of PD1.5 onto human kappa and IgG4 constant regions containing an S228P mutation (prevents Fab arm exchange with endogenous IgG4 antibodies).

-20°C,2°C to 8°C

12352203