Denosumab antibody | AbD26862_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-Denosumab Antibody (clone AbD26862_hIgG1, HCA282) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit®.
A microtiter plate was coated overnight with denosumab, human IgG2/kappa or human IgG1/lambda at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed Human Anti-Denosumab Antibody (clone AbD26862, here as his-tagged monovalent Fab fragment) titrated to the given concentrations in PBST, followed by detection with Mouse Anti-Penta Histidine Tag:HRP (MCA5995P) and QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of three measurements.
A microtiter plate was coated overnight with denosumab 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-Denosumab Antibodies, clone AbD26295_hIgG1 (HCA280, black line; KD = 1.8 nM), clone AbD26296_hIgG1 (HCA281, red line; KD = 7.1 nM) or clone AbD26862_hIgG1, HCA282 (blue line; KD = 0.8 nM). Detection was performed using HRP conjugated denosumab followed by QuantaBlu Fluorogenic Peroxidase Substrate. HRP conjugation of the denosumab used in this assay was performed using a LYNX Rapid HRP Conjugation Kit®. Data is shown as the mean of three measurements.
A microtiter plate was coated overnight with recombinant human RANKL at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, denosumab was added (0.3 μg/ml), spiked with increasing concentrations of each of three Human Anti-Denosumab Antibodies, clone AbD26295_hIgG1 (HCA280), and AbD26862_hIgG (HCA282), both in monovalent Fab format in this assay, and clone AbD26296 (HCA288). Detection of denosumab was performed using Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of three measurements.
Human anti Denosumab
Anti-denosumab antibody is a recombinant, inhibitory, anti-idiotypic antibody in IgG1 format for use in bioanalysis and immunogenicity testing of denosumab and biosimilars. It can be used as detection antibody in a PK bridging ELISA and as a reference standard in an ADA bridging ELISA.
- Product Type
- Monoclonal Antibody
- Clone
- AbD26862_hIgG1
- Isotype
- IgG1
| Product Code | Applications | Datasheet | MSDS | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|---|
| HCA282 | E | 0.1 mg |
Human Anti-Denosumab Antibody, clone AbD26862_hIgG1 is a paratope specific anti-idiotypic antibody that specifically recognizes the free human monoclonal antibody denosumab. The antibody does not recognize free RANKL (receptor activator of nuclear factor kappa-B ligand) or denosumab in complex with human RANKL and can be used to measure free denosumab and biosimilar products in bioanalytical assays.
Clone AbD26862_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.
Denosumab (Prolia, Xgeva) is a fully human monoclonal antibody (IgG2/kappa) for the treatment of osteoporosis, treatment-induced bone loss, bone metastases, multiple myeloma, and giant cell tumor of bone. The drug specifically binds to human RANKL a protein that acts as the primary signal to promote bone removal/resorption and prevents RANKL from activating its receptor, RANK, on the surface of osteoclasts and their precursors.
View a summary of all anti-denosumab antibodies.
Clone AbD26862_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.
Denosumab (Prolia, Xgeva) is a fully human monoclonal antibody (IgG2/kappa) for the treatment of osteoporosis, treatment-induced bone loss, bone metastases, multiple myeloma, and giant cell tumor of bone. The drug specifically binds to human RANKL a protein that acts as the primary signal to promote bone removal/resorption and prevents RANKL from activating its receptor, RANK, on the surface of osteoclasts and their precursors.
View a summary of all anti-denosumab 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 - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Denosumab
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=0.8 nM by real time, label-free molecular interaction analysis on immobilized denosumab
- Approx. Protein Concentrations
- IgG 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. Prolia and Xgeva are registered trademarks of Amgen Inc.
- 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 Denosumab 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
- This antibody is fully human and can be used as a reference standard in an ADA assay.
Protocol: ADA bridging ELISA
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Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | 50 ml | E IY | ||
| Human anti Denosumab | HCA280 | 0.1 mg | E | ||
| Human anti Denosumab | HCA281 | 0.1 mg | E |
