Golimumab antibody | AbD25705
A microtiter plate was coated over night 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 (clone AbD25705, HCA274) at a concentration of 2 μg/ml followed by Mouse anti His tag:HRP (MCA5995P) in HISPEC diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate.
A microtiter plate was coated over night with human TNFα, golimumab or human IgG1/kappa 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 (clone AbD25705, HCA274) titrated to the given concentrations in PBST, followed by Mouse anti His tag:HRP (MCA5995P) in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. Data for binding to human TNFα/golimumab are presented as the mean of three measurements.
A microtiter plate was coated over night with human TNFα at a concentration of 5 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 drug/target complex (clone AbD25705, HCA274) at a concentration of 2 μg /ml, followed by Mouse anti His tag antibody:HRP (MCA5995P) in HISPEC diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. Data is presented as the mean of three measurements
Human anti Golimumab (Drug/Target Complex)
- Product Type
- Monoclonal Antibody
- Clone
- AbD25705
- Isotype
- HuCAL Fab monovalent
- Specificity
- Golimumab
- Region
- DRUG/TARGET COMPLEX
Antibody Quality and Performance Guarantee
| Human anti golimumab (drug/target complex), clone AbD25705 is an affinity matured recombinant antibody in monovalent Fab format that specifically recognizes golimumab when in complex with its target, tumor necrosis factor alpha (TNFα). Clone AbD25705 does not recognize unbound golimumab nor free TNFα. The antibody can be used in a pharmacokinetic (PK) antigen capture format assay to measure golimumab that has been captured via immobilized human TNFα. It is ideal for bioanalytical assays to measure the level of drug or biosimilar in complex with the TNFα target. 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
- Source
- E.coli
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% Sodium Azide (NaN3)
- Immunogen
- Golimumab/TNFα complex
- Affinity
- The intrinsic affinity of the monovalent form of Human anti Golimumab (Drug/target complex) antibody, clone AbD25705 is KD = 6 nM as measured by real time, label free molecular interaction analysis on immobilized golimumab in complex with human TNFα.
- 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.
His-tag is a registered 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
- This product may be used in an indirect ELISA to detect the golimumab/TNFα complex or to detect golimumab bound to immobilized TNFα.
Protocol: PK antigen capture ELISA to measure bound drug exclusively.
| 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 | ||||||
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
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Log in | ||
| List Price | Your Price | ||||||
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| Description | Mouse anti Penta Histidine Tag: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): 178-190.
- 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.
HCA274
1611
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