Golimumab antibody | AbD25455
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-Golimumab Antibody, clone AbD25455 at a concentration of 2 μg/ml followed by Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. This antibody is affinity matured from parental antibody clone AbD20897 (HCA242). Both the affinity matured and parental clones exhibit some cross reactivity with omalizumab, the parental clone to a lesser extent.
A microtiter plate was coated overnight with golimumab, human TNFα, human IgG1/kappa, human IgG1/lambda or adalimumab at a concentration of 5 μg/ml. After washing and blocking with PBST + 5% BSA, the human TNFα/golimumab complex was formed by adding 2 μg/ml golimumab to the wells coated with human TNFα. Detection was performed using Human Anti-Golimumab Antibody, clone AbD25455 titrated to the given concentrations in PBST followed by Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data for golimumab are shown as the mean of three measurements.
A microtiter plate was coated overnight with human TNFα at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of golimumab (0.3 μg/ml) plus increasing concentrations of either Human Anti-Golimumab Antibody, clone AbD25418 (HCA287) or AbD25455 (HCA289) was added, both in monovalent Fab format in this assay. Free golimumab, still capable of binding to the human TNFα coated plate, was detected using Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Golimumab Antibody, clone AbD25455 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 golimumab. Detection was performed using Human Anti-Golimumab Antibody, clone AbD25451_hIgG1, (HCA290P) 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.
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E ResetHuman anti Golimumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD25455
- Isotype
- HuCAL Fab monovalent
- Specificity
- Golimumab
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| Human Anti-Golimumab Antibody, clone AbD25455 is an anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug golimumab. This antibody binds to both free golimumab and to golimumab bound to its target tumor necrosis factor alpha (TNFα). Clone AbD25455 does not inhibit the binding of golimumab to its target TNFα and can be used to measure total golimumab and biosimilar products in bioanalytical assays. Clone AbD25455 is an affinity-matured variant of clone AbD20897 (HCA242), with about 7-fold affinity improvement. A pair of anti golimumab 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 antibody in full immunoglobulin format, clone AbD25451_hIgG1 (HCA290), as the detection antibody. Golimumab, marketed under the brand name Simponi is a recombinant human IgG1/kappa monoclonal antibody approved for the treatment of rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis. This therapeutic antibody, directed against TNFα, acts by blocking the binding of TNFα to its receptors, resulting in a down-regulation of the inflammatory response associated with autoimmune diseases. View a summary of all anti-golimumab 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
- Golimumab
- Affinity
- The intrinsic affinity of the monovalent form of Human anti Golimumab antibody, clone AbD25455 is KD = 0.9 nM as measured by real time, label free molecular interaction analysis on immobilized Golimumab.
- 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.
Simponi is a trademark of Janssen Biotech, Inc. in the USA and Merck & Co, Inc. in Europe.
His-tag is a 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
- Human anti Golimumab antibody, clone AbD25455 can be used in a direct or indirect ELISA system or as capture antibody for golimumab in a bridging ELISA together with HCA290 (AbD25455_hIgG1) as the detection reagent.
Protocol: PK bridging ELISA to measure total drug.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
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| Description | Mouse anti Penta Histidine Tag:HRP | ||||||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK001P | CJ | 1 Conjugation For 400µg Antibody | Log in | |||
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| Description | LYNX Rapid HRP Antibody Conjugation Kit | ||||||
| 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 | ||||||
| Human anti Golimumab | HCA290 | E | 0.1 mg |
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| Description | Human anti Golimumab | ||||||
| Human anti Golimumab:HRP | HCA290P | E | 0.1 mg |
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| Description | Human anti Golimumab:HRP | ||||||
References for Golimumab antibody
-
Harth, S. et al. (2019) Generation by phage display and characterization of drug-target complex-specific antibodies for pharmacokinetic analysis of biotherapeutics.
MAbs. 11 (1): 178-90.
- Synonyms
- Simponi
- 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.
View more products with Golimumab specificity
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