Nivolumab (anti-PD-1)

Catalog No.A2002 Batch: A200205

Technical Data

CAS No.	946414-94-4
Formulation	PBS buffer, pH 7.2
Isotype	Human IgG4
Source	CHO cells
Storage (From the date of receipt)	Store the undiluted solution at 4°C in the dark to avoid freeze-thaw cycles
Purity	99.5%
Protein concentration	5mg/ml
Endotoxin Level	≤1 EU/mg

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

The cell binding ratio of ¹²⁵I-Nivolumab and ¹²⁵I-Nivolumab-DTPA to PD-1 is 13.78 ± 2.90% and 10.22 ± 2.62%, respectively. Conjugating of DTPA does not affect targeting of Nivolumab to PD-1 in vitro.^[1]

In vivo

In SPECT images, lesions with high ^99mTc-DTPA-nivolumab uptake and relatively clear background are shown at 6 h. Thereafter, the suspected intestinal thickening in Gd-free T1WI is observed at 2 h after the addition of Gd-DTPA-nivolumab.^[1]

Protocol (Only for Reference)

Cell Assay:	Splenocytes were isolated from the spleens obtained from humanized PD-1 over-expressing mice following standard protocol. ¹²⁵I labeled Nivolumab and Nivolumab-DTPA, respectively. To perform co-culture, ¹²⁵I-Nivolumab and ¹²⁵I-Nivolumab-DTPA (0.37 MBq/µg) were added to the freshly isolated splenocytes (1×10⁶ cells) grown in 6-well plates, respectively. After incubation for 12 h at 4°C, cells were collected after rinsing three times with PBS. ¹²⁵I radioactivity was determined in a gamma counter to evaluate their binding efficiency to PD-1.
Animal Study:	Animal Models: CRC mouse models (highly expressed human PD-1 antigen) Dosages: 11.1 MBq/10 μg Administration: i.v. Reference: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8883027/

Cell Assay:

Splenocytes were isolated from the spleens obtained from humanized PD-1 over-expressing mice following standard protocol. ¹²⁵I labeled Nivolumab and Nivolumab-DTPA, respectively. To perform co-culture, ¹²⁵I-Nivolumab and ¹²⁵I-Nivolumab-DTPA (0.37 MBq/µg) were added to the freshly isolated splenocytes (1×10⁶ cells) grown in 6-well plates, respectively. After incubation for 12 h at 4°C, cells were collected after rinsing three times with PBS. ¹²⁵I radioactivity was determined in a gamma counter to evaluate their binding efficiency to PD-1.

Animal Study:

Animal Models: CRC mouse models (highly expressed human PD-1 antigen)
Dosages: 11.1 MBq/10 μg
Administration: i.v.
Reference: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8883027/

References	[1] Li D, et al. Front Bioeng Biotechnol. 2022 Feb 14;10:839756.

References

[1] Li D, et al. Front Bioeng Biotechnol. 2022 Feb 14;10:839756.

Customer Product Validation

Data from [Data independently produced by , , Clin Cancer Res, 2019, doi:10.1158/1078-0432]

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).

Data from [Data independently produced by , , Ann Surg Oncol, 2019, 26(1):139-147]

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.

Selleck's Nivolumab (anti-PD-1) has been cited by 67 publications

Genomic and transcriptomic profiling of peripheral T cell lymphoma reveals distinct molecular and microenvironment subtypes [ Cell Rep Med, 2024, 5(2):101416]	PubMed: 38350451
NKG2A+CD8+ T cells infiltration determines immunosuppressive contexture and inferior response to immunotherapy in clear cell renal cell carcinoma [ J Immunother Cancer, 2024, 12(1)e008368]	PubMed: 38262706
[ Transl Oncol, 2024, ]	PubMed: 38412661
Tumor associated neutrophils governs tumor progression through an IL-10/STAT3/PD-L1 feedback signaling loop in lung cancer [ Transl Oncol, 2024, 40:101866]	PubMed: 38128466
Variable PD-1 glycosylation modulates the activity of immune checkpoint inhibitors [ Life Sci Alliance, 2024, 7(3)e202302368]	PubMed: 38176728
A murine model to evaluate immunotherapy effectiveness for human Fanconi anemia-mutated acute myeloid leukemia [ PLoS One, 2024, 19(1):e0292375]	PubMed: 38289944
Acquired resistance to CDK4/6 inhibition is associated with dysregulation of multiple pathways including cyclin D-CDK4/6-RB, EGFR/HER, AKT/mTOR and IFN [ Research Square, 2024, 10.21203/rs.3.rs-3782509/v1]	PubMed: none
Base editing screens map mutations affecting interferon-γ signaling in cancer [ Cancer Cell, 2023, S1535-6108(22)00595-5]	PubMed: 36669486
Low-dose carboplatin modifies the tumor microenvironment to augment CAR T cell efficacy in human prostate cancer models [ Nat Commun, 2023, 14(1):5346]	PubMed: 37660083
Skin basal cell carcinomas assemble a pro-tumorigenic spatially organized and self-propagating Trem2+ myeloid niche [ Nat Commun, 2023, 14(1):2685]	PubMed: 37164949

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS.

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