Pembrolizumab antibody | AbD30689_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 AbD30689_hIgG1 (HCA298) 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 AbD30689_hIgG1 (HCA298) 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 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 AbD30689_hIgG1 (HCA298). 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 AbD30689 (HCA296) was added; HCA296 is the same clone as HCA298, but in monovalent Fab format. 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.
Human anti Pembrolizumab
Human Anti-Pembrolizumab Antibody is a recombinant, inhibitory, anti-idiotypic antibody in IgG1 format for use in immunogenicity testing. It can be used as a calibrator in an anti-drug antibody assay.
- Product Type
- Monoclonal Antibody
- Clone
- AbD30689_hIgG1
- Isotype
- IgG1
| Product Code | Applications | Datasheet | MSDS | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|---|
| HCA298 | E | 0.1 mg |
Human Anti-Pembrolizumab Antibody, clone AbD30689_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 AbD30689_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.
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
Clone AbD30689_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.
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 Details
- 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 AbD30689_hIgG1 was measured as KD = 0.6 nM by real time, label free molecular interaction analysis on immobilized pembrolizumab.
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Shelf Life
- 12 months from date of despatch
More Information
- Acknowledgements
- Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany. Keytruda is a trademark of Merck & Co.
- 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.
- Regulatory
- For research purposes only
Applications of Pembrolizumab antibody
| 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 AbD30689_hIgG1 can be used in a direct or indirect ELISA or as a detection antibody for pembrolizumab. It may also be used as a surrogate positive control to generate a standard curve for an anti-drug antibody assay.
Protocol: anti-drug antibody assay
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Negative Isotype Controls Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Recombinant Human IgG1 Lambda | HCA049 | 0.1 mg | E |
Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | 50 ml | E IY | ||
| Human anti Pembrolizumab | HCA296 | 0.1 mg | E | ||
| Human anti Pembrolizumab | HCA297 | 0.1 mg | E | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | 3 Conjugations For 400µg Antibody | CJ | ||
| Mouse anti Human IgG (Fc) CH2 Domain:HRP | MCA647P | 0.2 mg | C* E |
