Natalizumab Antibody | AbD21375

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Human anti natalizumab 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 natalizumab (clone AbD21375) at a concentration of 2 μg/ml followed by Rat anti DYKDDDDK Tag:HRP (MCA4764P) in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate

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Human anti natalizumab specificity titration ELISA

A microtiter plate was coated over night with natalizumab, human IgG4/kappa, human IgG1/kappa, human IgG1/lambda, tocilizumab or omalizumab at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human anti natalizumab (clone AbD21375) titrated to the given concentrations in PBST, followed by an HRP conjugated anti-His tag antibody in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. Data is presented as the mean of three measurements

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Natalizumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti natalizumab (clone AbD21375) 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 natalizumab. Detection was performed using HRP-conjugated Human anti natalizumab (clone AbD21375) 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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Natalizumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti natalizumab (clone AbD21375) 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 natalizumab. Detection was performed using HRP-conjugated Human anti natalizumab (clone AbD21375_hIgG1, HCA249) 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 natalizumab inhibits the binding of natalizumab to alpha-4/beta-1 integrin

A microtiter plate was coated over night with alpha-4/beta-1 integrin at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of natalizumab (0.3 μg/ml) plus increasing concentrations of Human anti natalizumab (clone AbD21375) was added. Free natalizumab, still capable of binding to the alpha-4/beta-1 integrin 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 Natalizumab
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-natalizumab antibody is a recombinant, neutralizing anti-idiotypic antibody in monovalent Fab format; it is ideal for bioanalytical assay development to measure levels of natalizumab or biosimilars. It is recommended as capture antibody in PK bridging ELISA with HCA249/HCA249P.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD21375
    • Isotype
      HuCAL Fab monovalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA248Edatasheet pdfdatasheet pdf0.1 mg
      HCA248
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
      -
      • Human anti natalizumab, clone AbD21375 is a Type 1 anti-idiotypic antibody that specifically recognizes free natalizumab but not free human alpha-4/beta-1 integrin or the drug/integrin complex. Clone AbD21375 inhibits the binding of natalizumab to alpha-4/beta-1 integrin and can be used to measure free natalizumab levels in serum from patients. This clone is also suitable for pharmacokinetic (PK) assay development.

        Natalizumab, marketed under the brand name Tysabri®, is a humanized IgG4/kappa monoclonal antibody approved as a monotherapy for the treatment of relapsing forms of multiple sclerosis and for the treatment of Crohn’s disease. Natalizumab is also being evaluated for potential clinical use in the treatment of B-cell malignancies. This therapeutic antibody, directed against alpha-4 integrin (CD49d), binds to alpha-4/beta-1 integrin on the surface of circulating leucocytes. It is believed to act by blocking the interaction of alpha-4/beta-1 integrin with vascular cell adhesion molecule-1 (VCAM-1), preventing the adherence and transport of inflammatory immune cells across the intestinal lining and blood-brain barrier.

        Clone AbD21375 is one of a range of Type 1 (HCA249, HCA250), anti-idiotypic antibodies to natalizumab 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
        Natalizumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD = 2.1 nM by real time, label free molecular interaction analysis on immobilized natalizumab.
      • 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, a wholly owned subsidiary of Bio-Rad Laboratories Inc., Zeppelinstr. 4, 82178 Puchheim, Germany.

        Tysabri is a registered trademark Biogen Idec and Élan.
      • 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 a either a direct ELISA or indirect ELISA both as a capture reagent and as a detection reagent.
        Protocol: PK bridging ELISA to measure free drug
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Natalizumab Antibody Formats

      Formats Clone Applications Sizes available
      Natalizumab Antibody : Purified AbD21375 E 0.1 mg
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Rat anti DYKDDDDK Tag:HRPMCA4764P0.1 mgE, P, WB
            MCA4764P
            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 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
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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