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.
Human anti Golimumab
Anti-Golimumab Antibody is a recombinant, non-inhibitory, anti-idiotypic antibody in monovalent Fab format, suitable for bioanalytical assay development for golimumab and biosimilars. It is suitable as a capture antibody in a PK bridging ELISA to measure total drug, paired with HCA290.
- Product Type
- Monoclonal Antibody
- Clone
- AbD25455
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Datasheet | MSDS | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|---|
| HCA289 | E | 0.1 mg |
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
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 Details
- 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 monovalent intrinsic affinity of AbD25455 was measured as KD = 0.9 nM by real time, label free molecular interaction analysis on immobilized golimumab
- 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. Simponi is a registered trademark of Janssen Biotech, Inc. in the USA and Merck & Co, Inc. in Europe.
His-tag is a registered trademark of EMD Biosciences. - 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 Golimumab 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 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
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Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Human anti Golimumab | HCA290 | 0.1 mg | E | ||
| Human anti Golimumab:HRP | HCA290P | 0.1 mg | E | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK001P | 1 Conjugation For 400µg Antibody | CJ | ||
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | 125 µl | E WB | ||
| Mouse anti Human IgG (Fc) CH2 Domain:HRP | MCA647P | 0.2 mg | C* E |
