Adalimumab Antibody | AbD20350

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Human anti adalimumab 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 adalimumab (clone 20350) at a concentration of 2 μg/ml, followed by an HRP-conjugated mouse anti His tag antibody in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate

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Human anti adalimumab specificity titration ELISA

A microtiter plate was coated over night with various antigens. For TNFα/drug complex antigens, human TNFα was immobilized to the plate. After washing and blocking, adalimumab or infliximab was added. After further incubation and washing steps, detection was performed using Human anti adalimumab (clone 20350) titrated to the given concentrations in PBST, followed by an HRP-conjugated mouse anti His tag antibody in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate

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Adalimumab 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 adalimumab. Detection was performed using unlabeled (red line) or HRP-conjugated (black line) Human anti adalimumab (clone 20350) at a concentration of 2 µg/ml in PBST. Unlabeled anti adalimumab was detected by an HRP-conjugated mouse anti His tag antibody in HISPEC assay diluent (BUF049C) and QuantaBlu fluorogenic peroxidase substrate. HRP-conjugated anti adalimumab was detected directly with QuantaBlu substrate. HRP-conjugation of the antibody used in this assay was performed using a Lynx Rapid HRP Antibody Conjugation Kit™

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  • Human anti Adalimumab (Drug/Target Complex)
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    Anti-adalimumab drug-target complex antibody specifically recognizes the adalimumab-TNF alpha complex exclusively and detects adalimumab or biosimilars only when bound to TNF alpha. This affinity-matured antibody is in monovalent Fab format and is suitable for a PK antigen capture format assay.
    • Product Type
      Monoclonal Antibody
    • Clone
    • Isotype
      HuCAL Fab monovalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA231Edatasheet pdfdatasheet pdf0.1 mg
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      • Human anti Adalimumab (drug/target complex), clone AbD20350, is an affinity matured, Type 3 anti-idiotypic antibody that specifically recognizes adalimumab complexed with TNFα. It does not recognize the free drug or unbound TNFα. Clone AbD20350 can be used in assays to measure adalimumab that has been captured via immobilized TNFα, thereby avoiding the bridging assay format. It can also be used to measure the level of the drug/target complex and biosimilar products using bioanalytical assays.

        Adalimumab (brand name Humira®) is a human monoclonal antibody (IgG1/kappa) approved for the treatment of rheumatoid, psoriatic and juvenile idiopathic arthritis, ankylosing spondylitis, Crohn’s disease and ulcerative colitis. This therapeutic antibody is directed against the pro-inflammatory cytokine tumor necrosis factor alpha (TNFα), blocking its interaction with the p55 (TNFR1, CD120a) and p75 (TNFR2, CD120b) cell surface TNF receptors, resulting in a down-regulation of the abnormal inflammatory response that is characteristic of autoimmune disease.

        View a summary of all anti-adalimumab antibodies.
      • Intended Use
      • Product Form
        A monovalent human recombinant Fab (kappa light chain) selected from the HuCAL® phage display library, expressed in E. coli. This antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This product is supplied as a liquid.
      • Reconstitution
      • Preparation
        Metal chelate affinity chromatography
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD=3.2 nM by real time, label-free molecular interaction analysis on immobilized adalimumab/TNFα complex.
      • 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.
        Humira® is a registered trademark of Abbott Laboratories.
      • 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

      • 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 adalimumab/TNFα complex or to detect adalimumab 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 Adalimumab Antibody Formats

      Formats Clone Applications Sizes available
      Adalimumab Antibody : Purified AbD20350 E 0.1 mg
      • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse anti Histidine Tag:Alk.Phos.MCA1396A0.1 mgE, WB
        Goat anti Histidine Tag:BiotinAHP1656B0.1 mgE, WB
        Mouse anti Histidine Tag:BiotinMCA1396B0.1 mgE, WB
        Rabbit anti DYKDDDDK Tag:HRPAHP1074P0.1 mgE, WB
        Rat anti DYKDDDDK Tag:HRPMCA4764P0.1 mgE, P, WB
        Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            LYNX Rapid HRP Antibody Conjugation KitLNK002P3 Conjugations For 400µg AntibodyCJ
            Human anti Adalimumab (Drug/Target Complex)HCA2320.1 mgE
            Hispec Assay DiluentBUF049A50 mlIY
            Recombinant Human TNF AlphaPHP05150 µgE, FN, WB

            Recommended Positive Controls

              Histology Controls

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