Pembrolizumab antibody | AbD30685_hIgG1
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 HRP conjugated Human Anti-Pembrolizumab Antibody, clone AbD30685_hIgG1 (HCA297) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with pembrolizumab or human IgG4-Pro, human PD-1, nivolumab, natalizumab or trastuzumab at a concentration of 5 µg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HRP conjugated Human Anti-Pembrolizumab Antibody, clone AbD30685_hIgG1 (HCA297) titrated to the given concentrations 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-Pembrolizumab Antibody, clone AbD30689 (HCA296) 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 pembrolizumab. Detection was performed using HRP conjugated Human Anti-Pembrolizumab Antibody, clone AbD30685_hIgG1 (HCA297) 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 pembrolizumab 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 Human Anti-Pembrolizumab Antibody, clone AbD30685_hIgG1 (HCA297). Detection was performed using HRP conjugated pembrolizumab followed by QuantaBlu Fluorogenic Peroxidase Substrate. HRP conjugation of the pembrolizumab used in this assay was performed using a LYNX Rapid HRP Conjugation Kit (LNK002P). 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 pembrolizumab (0.3 µg/ml), spiked with increasing concentrations of Human Anti-Pembrolizumab Antibody, clone AbD30685 (HCA297) in the monovalent Fab format was added. Free pembrolizumab, 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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E ResetHuman anti Pembrolizumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD30685_hIgG1
- Isotype
- IgG1
- Specificity
- Pembrolizumab
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| Human Anti-Pembrolizumab Antibody, clone AbD30685_hIgG1 is a paratope specific, inhibitory anti-idiotypic antibody that specifically recognizes the free humanized monoclonal antibody pembrolizumab. The antibody does not recognize recombinant human programmed cell death 1 (PD-1) or pembrolizumab in complex with recombinant human PD-1 and can be used to measure free pembrolizumab levels in serum from patients. Clone AbD30685_hIgG1 is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a reference standard in an anti-drug antibody (ADA) assay. A pair of anti-pembrolizumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in full immunoglobulin format, is recommended as the detection antibody paired with clone AbD30689 (HCA296) in monovalent Fab format as the capture antibody. Pembrolizumab (Keytruda) is a humanized (IgG4/kappa) antibody with the heavy chain mutation S228P (IgG4-Pro). Pembrolizumab is used to treat inoperable or metastatic melanoma, metastatic non-small cell lung cancer (NSCLC), and as a second-line treatment for head and neck squamous cell carcinoma (HNSCC). For NSCLC, pembrolizumab is a first line treatment if the cancer overexpresses PDL1 and has no mutations in EGFR or ALK. Pembrolizumab is a checkpoint inhibitor and acts as an immunomodulator by blocking ligand activation of PD-1 receptor on T cells, thereby activating T cells to attack the cancer. It blocks PD-1 interaction with PD-1 ligand (PD-L1) and PD-1 ligand 2 (PD-L2). View a summary of all anti-pembrolizumab antibodies |
- Product Form
- Human IgG1 antibody (lambda light chain) selected from the HuCAL® phage display library and expressed in a human cell line. This antibody is supplied as a liquid.
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Pembrolizumab
- Affinity
- The monovalent intrinsic affinity of AbD30685_hIgG1 was measured as KD = 5 nM by real time, label free molecular interaction analysis on immobilized pembrolizumab.
- 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.
Keytruda is a trademark of Merck & Co.
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 AbD30685_hIgG1 can be used in a direct or indirect ELISA or as detection antibody for pembrolizumab in a bridging ELISA together with HCA296 as the capture reagent. Furthermore, it may be used as a calibrator for the set-up of an anti-drug antibody assay.
Protocol: PK bridging ELISA to measure free drug and anti-drug antibody assay
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Recombinant Human IgG1 Lambda Allotype G1m3 | HCA049 | E | 0.1 mg |
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| Description | Recombinant Human IgG1 Lambda Allotype G1m3 | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Human anti Pembrolizumab | HCA296 | E | 0.1 mg |
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| Description | Human anti Pembrolizumab | ||||||
| Mouse anti Human IgG (Fc) CH2 Domain:HRP | MCA647P | C* E | 0.2 mg |
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| Description | Mouse anti Human IgG (Fc) CH2 Domain:HRP | ||||||
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
| List Price | Your Price | ||||||
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| Description | Hispec Assay Diluent | ||||||
| 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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