Golimumab Antibody | AbD20897

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Human anti golimumab 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 (clone AbD20897) at a concentration of 2 µg/ml, followed by an HRP labelled anti His-tag antibody in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate

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Human anti golimumab 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 TNFα/golimumab complex was formed by adding 2 µg/ml golimumab to TNFα-coated wells. Detection was performed using Human anti golimumab (clone AbD20897) titrated to the given concentrations in PBST, followed by an HRP labelled anti His-tag antibody in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. Data for the binding of antibody to golimumab and TNFα/golimumab is presented as the mean of three measurements

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Golimumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti golimumab (clone AbD20710, HCA240) 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 HRP-conjugated Human anti golimumab (clone AbD20897) at a concentration of 2 µg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. HRP-conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit®. Data is presented as the mean of three measurements

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Human anti golimumab does not inhibit the binding of golimumab to TNFα

A microtiter plate was coated over night with 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 the non-inhibitory Type 2 Human anti golimumab (clone AbD20897) or the inhibitory Type 1 Human anti golimumab (clone AbD20710, HCA240) was added. Free golimumab, still capable of binding to the TNFα-coated plate, was detected using Mouse anti Human IgG (Fc) CH2 domain:HRP (MCA647P) and QuantaBlu™ fluorogenic peroxidase substrate. Data is presented as the mean of three measurements

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  • Human anti Golimumab
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-golimumab antibody is a recombinant, non-neutralizing anti-idiotypic antibody in monovalent Fab format, suitable for bioanalytical method development for golimumab and biosimilars. It is suitable in a PK bridging ELISA to measure total drug.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD20897
    • Isotype
      HuCAL Fab monovalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA242Edatasheet pdfdatasheet pdf0.1 mg
      HCA242
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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      • Human anti golimumab, clone AbD20897 is a Type 2 anti-idiotypic antibody that specifically recognizes free golimumab and the drug/tumor necrosis factor alpha (TNFα) complex. Clone AbD20897 does not inhibit the binding of golimumab to TNFα and can be used to measure the total golimumab levels in serum from patients, or for pharmacokinetic (PK) assay development.

        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.

        Clone AbD20897 is one of a range of Type 1 (HCA240, HCA241), Type 2 (HCA243) and Type 3 (HCA244, HCA245) anti-idiotypic antibodies to golimumab available from Bio-Rad and forms part of a wider and expanding portfolio of anti-idiotypic antibodies that have been developed for the detection of biotherapeutics.
      • Intended Use
      • Product Form
        A monovalent human recombinant Fab 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 AbD20897 was measured as KD = 7 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 6,300,064, 6,696,248, 6,708,484, 6,753,136, European Patent 0,859,841 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany, a wholly owned subsidiary of Bio-Rad Laboratories Inc.

        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 AbD20897 can be used in an indirect ELISA system or, following HRP-conjugation, as a detection reagent for PK assay development together with clone AbD20710 (HCA240) as the capture reagent.
        Protocol: PK bridging ELISA to measure free drug
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Golimumab Antibody Formats

      Formats Clone Applications Sizes available
      Golimumab Antibody : Purified AbD20897 E 0.1 mg
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
        MCA5995P

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            LYNX Rapid HRP Antibody Conjugation KitLNK005P1 Conjugation For 20mg AntibodyCJ
            LNK005P
            LYNX Rapid HRP Antibody Conjugation KitLNK001P1 Conjugation For 400µg AntibodyCJ
            LNK001P
            LYNX Rapid HRP Antibody Conjugation KitLNK003P1 Conjugation For 4mg AntibodyCJ
            LNK003P
            LYNX Rapid HRP Antibody Conjugation KitLNK006P3 Conjugations For 40µg AntibodyCJ
            LNK006P
            LYNX Rapid HRP Antibody Conjugation KitLNK002P3 Conjugations For 400µg AntibodyCJ
            LNK002P
            LYNX Rapid HRP Antibody Conjugation KitLNK004P5 Conjugations For 4mg AntibodyCJ
            LNK004P
            Human anti GolimumabHCA2400.1 mgE
            HCA240
            Human anti GolimumabHCA2430.1 mgE
            HCA243
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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