Golimumab Antibody | AbD25705

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Human anti golimumab (drug/target complex) specificity ELISA

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.

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Human anti golimumab (drug/target complex) specificity titration ELISA

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.

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Golimumab antigen capture ELISA for PK assay development

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

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  • Human anti Golimumab (Drug/Target Complex)
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-golimumab drug-target complex antibody specifically recognizes the golimumab-TNF alpha complex and detects golimumab or biosimilars only when bound to TNF alpha. This antibody is in monovalent Fab format and is suitable for a PK antigen capture format assay to detect bound drug.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD25705
    • Isotype
      HuCAL Fab monovalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA274Edatasheet pdfdatasheet pdf0.1 mg
      HCA274
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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      • 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
      • Intended Use
      • 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.
      • Reconstitution
      • Preparation
        Metal chelate affinity chromatography
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Golimumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of AbD25705 was measured as KD = 6 nM 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
      • Reagents In The Kit
      • Preparing The Antibody
      • Test Principle
      • Buffer Solution
        Phosphate buffered saline
      • 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
      • 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.
        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
      • 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 NameYesNoMin DilutionMax 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
      • Recommended Protocol
      • 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
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Golimumab Antibody Formats

      Formats Clone Applications Sizes available
      Golimumab Antibody : Purified AbD25705 E 0.1 mg
      • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
            MCA5995P
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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