Trastuzumab Antibody | AbD18018_hIgG4Pro

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Trastuzumab bridging ELISA for Pharmacokinetic (PK) assay development, detection antibody in IgG4 isotype

A microtiter plate was coated over night with Human anti trastuzumab, clone AbD18141 (HCA169) at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, 10% human serum spiked with increasing concentrations of trastuzumab was added. Detection was performed using either HRP conjugated Human anti trastuzumab, clone AbD18018_hIgG4Pro (HCA270) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) (black), or clone AbD18018_hIgG4Pro at 2 μg/ml followed by Mouse anti-human IgG4:HRP (MCA2098P) at a 1:10,000 dilution (green) in HISPEC assay diluent; QuantaBlu™ fluorogenic peroxidase substrate was used

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Human anti trastuzumab (IgG4 isotype) 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% milk powder, detection was performed using Human anti trastuzumab (clone AbD18018_hIgG4Pro) (blue bars) or Human anti trastuzumab (clone AbD18018_hIgG1) (red bars) at a concentration of 2 μg/ml, followed by anti human IgG4:HRP (MCA2098P) at a 1:25,600 dilution in HISPEC assay diluent (BUF049A) and QuantaBlu&trade fluorogenic peroxidase substrate

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  • Human anti Trastuzumab
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    Anti-trastuzumab antibody is a recombinant, inhibitory, high affinity anti-idiotypic antibody in fully human immunoglobulin format (IgG4 isotype) for bioanalytical method development. It is recommended as the detection antibody in a PK bridging ELISA.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD18018_hIgG4Pro
    • Isotype
      IgG4
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA270Edatasheet pdfdatasheet pdf0.1 mg
      HCA270
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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      • Human anti trastuzumab, clone AbD18018_hIgG4Pro, is a paratope specific, high affinity, anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug trastuzumab and inhibits it binding to its target. It is a fully human recombinant monoclonal antibody with IgG4 isotype. The antibody can be used to measure the levels of trastuzumab and biosimilar products in bioanalytical assays.

        Anti-trastuzumab antibody clone AbD18018 has been converted to human IgG4 isotype to offer assay developers increased flexibility in assay design. When used as an ELISA detection antibody, a secondary anti-human IgG4 antibody is required, which can be labeled according to the requirements of the user's preferred assay platform.

        The antibody can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug in preclinical research and clinical trials using patient sera. Antibody clones AbD16712 (HCA166) or AbD18141 (HCA169), both monovalent Fab format, are recommended as the capture antibody. Detection can be performed using clone AbD18018_hIgG4Pro and an HRP conjugated anti-human IgG4 secondary antibody (MCA2098P).

        This clone is also available as an IgG1 isotype in both a purified unconjugated format (HCA177) and directly conjugated to HRP (HCA177P).

        Trastuzumab, also known as Herceptin®, is a drug used in the treatment of HER2 postive breast cancer and other HER2 over-expressing cancers including HER2-positive metastatic cancers of the gastrointestinal tract. Trastuzumab binds to the HER2 (or c-erbB2) proto-oncogene, an EGF receptor-like protein found on 20-30% of breast cancer cells. The binding leads to antibody mediated (complement mediated) killing of the HER2 positive cells.

        View a summary of all anti-trastuzumab antibodies
      • Intended Use
      • Product Form
        Human IgG4 antibody selected from the HuCAL® phage display library and expressed in a human cell line - liquid
      • Reconstitution
      • Preparation
        Purified IgG prepared by affinity chromatography on Protein A
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Trastuzumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD=0.02 nM by real time, label-free molecular interaction analysis on immobilized trastuzumab.
      • Approx. Protein Concentrations
        IgG 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.
      • 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 as a detection reagent in a sandwich ELISA together with an HRP conjugated anti human IgG4 secondary antibody (MCA2098P) and HCA169 as the capture reagent.

        Protocol: In preparation.
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Trastuzumab Antibody Formats

      Formats Clone Applications Sizes available
      Trastuzumab Antibody : Purified AbD18018_hIgG4Pro E 0.1 mg
      • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse anti Human IgG4:HRPMCA2098P0.2 mgC, E, P
        MCA2098P

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Human anti Trastuzumab:HRPHCA177P0.1 mgE
            HCA177P
            Human anti TrastuzumabHCA1660.1 mgE
            HCA166
            Human anti TrastuzumabHCA1690.1 mgE
            HCA169
            Human anti TrastuzumabHCA1770.1 mgE
            HCA177
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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