Nivolumab (anti-PD-1)

Catalog No.A2002 Synonyms: BMS-936558, ONO-4538, MDX-1106

For research use only. Not for use in humans.

Nivolumab (anti-PD-1) is a genetically engineered, fully human immunoglobulin (Ig) G4 monoclonal antibody directed against the negative immunoregulatory human cell surface receptor programmed death-1 (PD-1,PCD-1) with immune checkpoint inhibitory and antineoplastic activities. MW : 143.597 KD.

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Cited by 6 Publications

2 Customer Reviews

  • Apoptosis rates of Jurkat cells co-cultured for 72 h with H1975, A549 cells or H1975 with 10 μg/mL nivolumab were analyzed by Annexin V-FITC/PI assay (Left) and quantified (Right).

    Clin Cancer Res, 2019, doi:10.1158/1078-0432. Nivolumab (anti-PD-1) purchased from Selleck.

    Preliminary immunotherapy drug screening using immune checkpoint inhibitors pembrolizumab and nivolumab in LGA organoids immune-enhanced with cells from a lymph node from the same patient. Mitochondrial metabolism was assessed at (c) 24 h and (d) 96 h after administration of the immunotherapy agents. Statistical significance: **p<0.05 between tumor cell-only and immune-enhanced organoids; p<0.05 between drug treatment and control of the same group.

    Ann Surg Oncol, 2019, 26(1):139-147. Nivolumab (anti-PD-1) purchased from Selleck.

Quality Control

Choose Selective PD-1/PD-L1 Inhibitors

Biological Activity

Description Nivolumab (anti-PD-1) is a genetically engineered, fully human immunoglobulin (Ig) G4 monoclonal antibody directed against the negative immunoregulatory human cell surface receptor programmed death-1 (PD-1,PCD-1) with immune checkpoint inhibitory and antineoplastic activities. MW : 143.597 KD.
Targets
PD-1/PD-L1 interaction [2]
(Cell-free assay)
PD-1/PD-L2 interaction [2]
(Cell-free assay)
2.52 nM 2.59 nM
In vitro

Nivolumab (anti-human PD-1) binds PD-1 with high affinity (KD 2.6 nmol/l by Scatchard analysis to polyclonally activated human T cells) and blocks its interactions with both B7-H1 and B7-DC[1]. It effectively inhibits the interaction between PD-1 and its ligands. In vitro assays demonstrated the ability of nivolumab to potently enhance T-cell responses and cytokine production in the mixed lymphocyte reaction and superantigen or cytomegalovirus stimulation assays. Nivolumab inhibits the interaction between PD-1 and its ligands, PD-L1 and PD-L2, with IC50 values of 2.52 and 2.59 nmol/L, respectively, as shown by surface plasmon resonance. In a study using FACS to evaluate ligand binding to PD-1 expressed on CHO cells, the IC50 values for nivolumab-mediated inhibition of PD-1 binding to PD-L1 or PD-L2 were similar (1.04 and 0.97 nmol/L, respectively). Nivolumab binds specifically to PD-1 and not to other immunoglobulin superfamily proteins, such as CD28, CTLA-4, ICOS, and BTLA. Nivolumab can, at very low concentrations (∼1.5 ng/mL), enhance T-cell reactivity in the presence of a T-cell receptor stimulus. However, nivolumab had no stimulatory effect in the absence of antigen or T-cell receptor stimulus. Specifically, there was no significant release of inflammatory cytokines, including IFNγ, TNFα, IL-2, IL-4, IL-6, or IL-10, from unstimulated whole blood after coincubation with nivolumab. Nivolumab does not cause nonspecific lymphocyte activation[2].

In vivo Nivolumab (anti-human PD-1) is well tolerated, dose-limiting toxicities (DLTs) were not reached and the maximum tolerable dose (MTD) was not defined in patients with advanced stage solid tumors. The measured half-life of nivolumab was 12-20 days, the pharmacodynamic effects of PD-1 receptor occupancy was even more prolonged at 85 days, indicating the biological durability of this high-affinity mAb[1]. In monkeys, serum nivolumab has a relatively slow clearance with limited extra vascular distribution, as demonstrated by a Vss value consistent with plasma volume. Mean apparent terminal elimination half-life estimates for males and females at 1 mg/kg were similar (124 and 139 hours, respectively), and the mean half-life estimate for males at 10 mg/kg was 261 hours. Although nivolumab seems to lack toxicity in monkeys, toxicities have been observed in human clinical trials. In a phase I trial, nivolumab had a favorable safety profile. Adverse events were generally similar to those observed with ipilimumab, although with lower incidence and of less severity, and comprised gastrointestinal, endocrine, and skin toxicities, and pulmonary inflammation. Interestingly, pneumonitis has been observed in PD-1-deficient mice bred onto the MRL genetic background, but not in PD-1-deficient mice with other genetic backgrounds[2].

Protocol

Cell Research:
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  • Objective: Antigen-specific recall response in vitro
    Cells: PBMCs
    Concentrations: 0-10 μM
    Incubation Time: 4 days
    Method: In a cytomegalovirus (CMV)-restimulation assay, 2 × 105 PBMCs from a CMV-positive donor (Astarte) were stimulated using lysate of CMV-infected cells (Astarte), with serial dilutions of nivolumab added at the initiation of the assay. After 4 days, supernatants were assayed for IFNγ.
    Reference: http://cancerimmunolres.aacrjournals.org/content/2/9/846

    Objective: Mixed lymphocyte response (MLR) assays
    Cells: Monocyte-derived DC and CD4+ T cells
    Concentrations: 50 mg/mL to 5 ng/mL
    Incubation Time: 6 days
    Method: Mixed lymphocyte response assays were performed by co-culturing 1×105 cells CD4+ T cells with allogeneic monocyte-derived dendritic cells (DC) at a ratio of 10:1 (T:DC) in flat-bottom 96-well microtiter plates. CD4+ T cells and DC were incubated for 6 days in the presence or absence of nivolumab titrated 1:10 from 50 mg/mL to 5 ng/mL along with ipilimumab at 0, 5, or 50 μg/mL. Culture supernatants were harvested on day 5 for ELISA analysis of IFN-γ secretion.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/27610613

    Nivolumab can apply to humanized mice (eg Rag2-/-IL2Rgnull mice), peripheral blood and other related assays (Only for Reference)

Animal Research:
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  • Objective: pharmacokinetic (PK) study
    Animal Models: Cynomolgus monkeys (Macaca fascicularis)
    Formulation: Saline
    Dosages: 3 mg/kg or 10 mg/kg
    Administration: i.v.
    Reference: http://cancerimmunolres.aacrjournals.org/content/2/9/846

    Objective: To determine that nivolumab and urelumab enhance antitumor activity in mice model
    Animal Models: Rag2-/-IL2Rgnull mice (humanized mice) were injected with 1×107 human PBMCs intraperitoneally (i.p.) and a total of 3.5×106 HT29 human colon carcinoma cells subcutaneously (s.c.)
    Formulation: saline
    Dosages: 200 μg per injection
    Administration: i.v.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/26113085

    Nivolumab can apply to humanized mice (eg Rag2-/-IL2Rgnull mice), peripheral blood and other related assays (Only for Reference)

Product Details

Formulation PBS buffer, pH 7.2
Isotype Human IgG4
Source CHO cells
Storage Store the undiluted solution at 4 °C in the dark and avoid freezing

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Frequently Asked Questions

  • Question 1:

    How to store the antibody?

  • Answer:

    Store the undiluted solution at 4 °C in the dark. Freezing antibodies can result in a loss of activity caused by the freezing/thawing process. Diluting antibodies to working concentrations and storing at 4°C for more than a day should be avoided. Additionally, make sure to keep the antibody sterile. Under these storage conditions, your antibodies should remain active for up to one year and oftentimes longer.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID