Nivolumab antibody | AbD30255
A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Nivolumab Antibody, clone AbD30255 (HCA299) at a concentration of 2 μg/ml and an HRP labeled anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with nivolumab or human IgG4-Pro, human PD-1, pembrolizumab, natalizumab or trastuzumab at a concentration of 5 µg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Nivolumab Antibody, clone AbD30255 (HCA299) titrated to the given concentrations in PBST followed by an HRP labeled anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Nivolumab Antibody, clone AbD30255 (HCA299) at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of nivolumab. Detection was performed using HRP conjugated Human Anti-Nivolumab Antibody (HCA301) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with recombinant human PD-1 at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of nivolumab (0.3 µg/ml), spiked with increasing concentrations of Human Anti-Nivolumab Antibody, clone AbD30255 (HCA299) was added. Free nivolumab, still capable of binding to the human PD-1 coated plate, was detected using a Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
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WB E ResetHuman anti Nivolumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD30255
- Isotype
- HuCAL Fab monovalent
- Specificity
- Nivolumab
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| Human Anti-Nivolumab Antibody, clone AbD30255 is a paratope specific, inhibitory anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug nivolumab. The antibody does not recognize recombinant human programmed cell death 1 (PD-1) or nivolumab in complex with recombinant human PD-1 and can be used to measure free nivolumab levels in serum from patients. A pair of anti-nivolumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in monovalent Fab format, is recommended as the capture antibody, paired with an HRP conjugated Anti-Nivolumab Antibody in full immunoglobulin format, clone AbD30258_hIgG1 (HCA301) as the detection antibody. Nivolumab (Opdivo) is a fully human antibody (IgG4/kappa), with the heavy chain mutation S228P (IgG4-Pro). It is used as a first line treatment for inoperable or metastatic melanoma in combination with ipilimumab if the cancer does not have a mutation in BRAF. It is also approved for treatment of advanced non-small cell lung cancer, advanced renal cell carcinoma, classical Hodgkin's lymphoma and advanced head and neck squamous cell carcinoma. Nivolumab is a checkpoint inhibitor and acts as an immuno-modulator by blocking ligand activation of PD-1 receptor on T cells, thereby activating T cells to attack the cancer. Nivolumab blocks PD-1 interaction with PD-1 ligand (PD-L1) and PD-1 ligand 2 (PD-L2). View a summary of all anti-nivolumab antibodies |
- Product Form
- A monovalent human recombinant Fab (lambda light chain) selected from the HuCAL® phage display library, expressed in E. coli. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This antibody is supplied as a liquid.
- Preparation
- Metal chelate affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Nivolumab
- Affinity
- The monovalent intrinsic affinity of AbD30255 was measured as KD = 0.5 nM by real time, label free molecular interaction analysis on immobilized nivolumab
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
Opdivo is a trademark of Bristol-Myers Squibb Company.
His-tag is a registered trademark of EMD Biosciences.
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Technical Advice
- Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- ELISA
- Clone AbD30255 can be used in a direct or indirect ELISA system or as capture antibody for nivolumab in a bridging ELISA together with HCA301 (AbD30258_hIgG1) as the detection reagent.
Protocol: PK bridging ELISA to measure free drug
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Rat anti DYKDDDDK Tag | MCA4764 | E F IF IP P WB | 0.2 mg |
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| Description | Rat anti DYKDDDDK Tag | ||||||
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
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| Description | Mouse anti Penta Histidine Tag:HRP | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| HuCAL Fab-FH Negative Control | HCA045 | E | 0.1 mg |
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| Description | HuCAL Fab-FH Negative Control | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
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| Description | Hispec Assay Diluent | ||||||
| Human anti Nivolumab | HCA301 | E | 0.1 mg |
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| Description | Human anti Nivolumab | ||||||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | Log in | |||
| List Price | Your Price | ||||||
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| Description | LYNX Rapid HRP Antibody Conjugation Kit | ||||||
- Licensed Use
- For in vitro research purposes and for commercial applications for the provision of in vitro testing services to support preclinical and clinical drug development. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.
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