Eculizumab antibody | AbD32107_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-Eculizumab Antibody, clone AbD32107_hIgG1 (HCA313), 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 eculizumab, human IgG4 kappa or human IgG2 kappa at a concentration of 5 μg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using HRP conjugated Human Anti-Eculizumab Antibody, clone AbD32107_hIgG1 (HCA313), titrated to the given concentrations in HISPEC Assay Diluent (BUF049A), and QuantaBlu Fluorogenic Peroxidase Substrate. Data for eculizumab are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Eculizumab Antibody, clone AbD32311 (HCA312), at a concentration of 1 μg/ml. After washing and blocking with PBST + 5% BSA, 10% fetal calf serum was added spiked with increasing concentrations of eculizumab. Detection was performed using HRP conjugated Human Anti-Eculizumab Antibody, clone AbD32107_hIgG1 (HCA313P), 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 eculizumab at a concentration of 1 μg/ml. After washing and blocking with PBST + 5% BSA, PBST with 10% fetal calf serum was added, spiked with increasing concentrations of Human Anti-Eculizumab Antibody, clone AbD32107_hIgG1 (HCA313, black), clone AbD32311_hIgG1 (HCA314, red), or clone AbD32109_hIgG1 (HCA315, blue). Detection was performed using HRP conjugated eculizumab followed by QuantaBlu Fluorogenic Peroxidase Substrate. HRP conjugation of the eculizumab 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 human complement C5 at a concentration of 1 μg/ml. After washing and blocking with PBST + 5% BSA, a pre-incubated mixture of eculizumab (0.3 μg/ml), spiked with increasing concentrations of Human Anti-Eculizumab Antibody, clone AbD32107 (HCA313) in the monovalent format was added. Free eculizumab still capable of binding to the human complement C5 coated plate, was detected using 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 Eculizumab:HRP
- Product Type
- Monoclonal Antibody
- Clone
- AbD32107_hIgG1
- Isotype
- IgG1
- Specificity
- Eculizumab
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| Human Anti-Eculizumab Antibody, clone AbD32107_hIgG1 is a paratope specific, inhibitory anti-idiotypic antibody that specifically recognizes the free humanized monoclonal antibody eculizumab. It does not recognize free human complement C5 or eculizumab in complex with C5. The antibody can be used to measure free eculizumab or eculizumab biosimilars in patient samples. Clone AbD32107_hIgG1 is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a surrogate positive control or calibrator in an anti-drug antibody (ADA) assay. A pair of anti-eculizumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody is recommended as the detection reagent, paired with Anti-Eculizumab Antibody clone AbD32311 (HCA312) in monovalent Fab format as the capture antibody. Eculizumab (Soliris) is a humanized IgG2/4 kappa antibody. The heavy chains are comprised of human IgG2 sequences in CH1, the hinge and the adjacent portion CH2, and human IgG4 sequences in the remaining part of CH2 and CH3. The drug is used to treat paroxysmal nocturnal hemoglobinuria (PNH), a rare acquired life-threatening disease of the blood characterized by destruction of red blood cells by the complement system. It also has approval as an orphan drug for treatment of atypical hemolytic uremic syndrome (aHUS). Eculizumab specifically binds to the terminal complement component 5 (C5) and inhibits the cleavage of C5 to C5a and C5b by the C5 convertase, which prevents the generation of the terminal complement complex C5b-9. Both C5a and C5b-9 cause the terminal complement-mediated events that are characteristic of PNH and aHUS. A 1:1 molar ratio of eculizumab to C5 is sufficient for C5 inhibition. View a summary of all anti-eculizumab antibodies |
- Product Form
- Human IgG1 antibody (kappa light chain) selected from the HuCAL® phage display library and expressed in a human cell line. Conjugated to horseradish peroxidase (HRP) - liquid.
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Eculizumab
- Affinity
- The monovalent intrinsic affinity of AbD32107_hIgG1 was measured as KD = 0.6 nM by real time, label free molecular interaction analysis on immobilized eculizumab.
- Approx. Protein Concentrations
- Antibody concentration 0.1 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.
Soliris is a trademark of Alexion Pharmaceuticals, Inc.
This product is shipped frozen.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
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 AbD32107_hIgG1 can be used in a direct ELISA or as the detection antibody for eculizumab in a bridging ELISA together with HCA312 as the capture reagent.
Protocol: PK bridging ELISA to measure free drug
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
| List Price | Your Price | ||||||
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| Description | Hispec Assay Diluent | ||||||
| Human anti Eculizumab | HCA312 | E | 0.1 mg |
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| List Price | Your Price | ||||||
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| Description | Human anti Eculizumab | ||||||
| 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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