Nivolumab (anti-PD-1)

目录号:A2002 别名: BMS-936558, ONO-4538, MDX-1106, Opdivo 中文名称:纳武单抗

For research use only.
Not for use in humans.

Nivolumab (anti-PD-1)是一种通过基因工程改造的、人类免疫球蛋白(Ig)G4单克隆抗体,靶向具有负向免疫调节功能的人类细胞表面受体程序性死亡-1(PD-1, PCD-1);也是一种抑制剂,具有免疫检查点抑制活性和抗肿瘤活性;MW:143.597 KD。

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客户使用Selleck生产的Nivolumab (anti-PD-1)发表文献19篇:

客户使用该产品的2个实验数据:

  • 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.

质量管理及产品安全说明书

PD-1/PD-L1抑制剂选择性比较

生物活性

产品描述 Nivolumab (anti-PD-1)是一种通过基因工程改造的、人类免疫球蛋白(Ig)G4单克隆抗体,靶向具有负向免疫调节功能的人类细胞表面受体程序性死亡-1(PD-1, PCD-1);也是一种抑制剂,具有免疫检查点抑制活性和抗肿瘤活性;MW:143.597 KD。
靶点
PD-1/PD-L1 interaction [2]
(Cell-free assay)
PD-1/PD-L2 interaction [2]
(Cell-free assay)
2.52 nM 2.59 nM
体外研究

Nivolumab (anti-human PD-1)与PD-1以高亲和力结合(Kd=2.6 nM),阻止其与B7-H1和B7-DC的相互作用[1]。它能有效地抑制PD-1和其配体的相互作用,体外试验证明nivolumab能有效增强T细胞反应、在混合淋巴细胞反应、超级抗原或巨细胞病毒刺激化验中增强细胞因子的产生。应用FACS来检测CHO细胞中PD-1的配体结合情况的实验中,nivolumab抑制PD-1与PD-L1或PD-L2结合的效力相似(IC50值相似,分别为1.04 nM和0.97 nM)。Nivolumab与PD-1特异性结合,而不与其他免疫球蛋白超家族蛋白如CD28、CTLA-4、ICOS和BTLA作用。在低浓度下(∼1.5 ng/mL),Nivolumab在存在T细胞受体刺激的情况下,可增强T细胞反应活性。然而,在缺少抗原或T细胞受体刺激的情况下,nivolumab没有刺激效果。在未受刺激的全血中,Nivolumab不会引起炎性因子(包括IFNγ, TNFα, IL-2, IL-4, IL-6或IL-10)的显著释放。Nivolumab也不会造成非特异性淋巴细胞的激活[2]

体内研究 Nivolumab (anti-human PD-1)在患者中耐受性良好,在具有晚期实体肿瘤的患者中,没有达到剂量限制性毒性,还没有确定最大耐受量。目前检测到的其在体内的半衰期为12-20天,它占据PD-1受体的药代动力学效果甚至可延长至85天。所以Nivolumab具有生物持久性[1]。在猴子中,血清中nivolumab具有相对较慢的清除率和有限的血管外分布。给药浓度为1 mg/kg时,在男性和女性中Nivolumab的Mean apparent terminal elimination half-life类似,分别为124和139小时。给药浓度为10 mg/kg时,nivolumab在男性体内的平均半衰期为261小时。尽管在猴子中nivolumab看似没有毒性作用,但在人类临床试验中观察到了其毒性作用。在临床I期试验中,nivolumab安全性良好。副作用与ipilimumab类似,尽管发生率较低,严重性较低,副作用包括胃肠道的、内分泌和皮肤毒性以及肺部炎症。肺炎只在具有PD-1缺陷的MRL遗传背景的小鼠中出现,而没有在其他遗传背景的PD-1缺陷型小鼠中观察到[2]

推荐的实验操作(此推荐来自于公开的文献所以Selleck并不保证其有效性)

细胞实验:
- 合并
  • 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)

动物实验:
- 合并
  • 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)

抗体信息

CAS No. 946414-94-4
配制 PBS buffer, pH 7.2
同型 Human IgG4
来源 CHO cells
储存条件 Store the undiluted solution at 4°C in the dark to avoid freeze-thaw cycles

技术支持

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常见问题及建议解决方法

  • 问题1:

    How to store the antibody?

  • 回答:

    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