Golimumab Antibody | AbD25455

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Human Anti-Golimumab Antibody specificity ELISA

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.

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Human Anti-Golimumab Antibody specificity titration ELISA

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.

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Human Anti-Golimumab Antibody inhibition ELISA

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.

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Human Anti-Golimumab Antibody bridging ELISA for pharmacokinetic (PK) assay development

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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  • Human anti Golimumab
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    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
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA289Edatasheet pdfdatasheet pdf0.1 mg
      HCA289
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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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
      • 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 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
      • 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. Simponi is a registered trademark of Janssen Biotech, Inc. in the USA and Merck & Co, Inc. in Europe
      • 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
        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
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Golimumab Antibody Formats

      Formats Clone Applications Sizes available
      Golimumab Antibody : Purified AbD25455 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
            Human anti Golimumab:HRPHCA290P0.1 mgE
            HCA290P
            Mouse anti Human IgG (Fc) CH2 Domain:HRPMCA647P0.2 mgC *, E
            MCA647P
            Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
            MCA5995P
            LYNX Rapid HRP Antibody Conjugation KitLNK001P1 Conjugation For 400µg AntibodyCJ
            LNK001P
            Human anti GolimumabHCA2900.1 mgE
            HCA290

            Recommended Positive Controls

              Histology Controls

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