Infliximab Antibody | AbD19376

//www.bio-rad-antibodies.com/infliximab-antibody-abd19376-hca214.html
Infliximab Antibody | AbD19376 gallery image 1

HCA214 Specificity ELISA

To confirm antibody specificity, a microtiter plate was coated overnight with various antigens at a concentration of 5µg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HCA214 at 2µg/ml, followed by Mouse anti Histidine-tag:HRP in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate

Enlarge
Infliximab Antibody | AbD19376 gallery image 2

HCA214 Specificity Titration ELISA

A microtiter plate was coated overnight with Human IgG1/kappa, Human IgG1/lambda or Infliximab at a concentration of 5µg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HCA214 titrated to the given concentrations in PBST+10% human serum, followed by Mouse anti Histidine-tag:HRP in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate

Enlarge
Infliximab Antibody | AbD19376 gallery image 3

HCA214 Sensitivity ELISA

To assess antibody sensitivity, a microtiter plate was coated overnight with HCA214 at 5µg/ml. After washing and blocking with PBST+5% BSA, Infliximab was added at the given concentrations. Detection was performed using Mouse anti Human IgG (Fc) CH2 domain:HRP (MCA647P) in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate

Enlarge
Infliximab Antibody | AbD19376 gallery image 4

Infliximab Inhibition ELISA

HCA214 does not inhibit the binding of Infliximab to TNF-α

To assess the ability of HCA214 to inhibit the binding of Infliximab to its target molecule, a microtiter plate was coated overnight with TNF-α at a concentration of 1µg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of HCA214 titrated into a constant amount of Infliximab (IFX, 0.3µg/ml) was added. Free Infliximab, still capable of binding to the plate, was detected using Mouse anti Human IgG (Fc) CH2 domain:HRP (MCA647P)

Enlarge
Infliximab Antibody | AbD19376 gallery image 5

Infliximab Inhibition ELISA

Infliximab bound to TNF-α still binds to HCA214

HCA214 was coated overnight onto a microtiter plate was coated overnight at a concentration of 5µg/ml. After washing and blocking with PBST+5% BSA, Infliximab alone (green triangles), with 0.5 fold molar excess of TNF-α (red circles) or with 5.0 fold molar excess of TNF-α (black squares) was added. Free or TNF-α bound Infliximab binding to HCA214 was detected using Mouse anti Human IgG (Fc) CH2 domain:HRP (MCA647P)

Enlarge
Infliximab Antibody | AbD19376 gallery image 6

Infliximab Bridging ELISA for Pharmacokinetic (PK) Assay Development

A microtiter plate was coated overnight with HCA214 at a concentration of 5µg/ml (black circles) or 1µg/ml (blue squares). After washing and blocking with PBST+5% BSA, Infliximab spiked in 10% human serum was added at the given concentrations. Detection was performed using HRP-conjugated HCA213 (black circles) or HCA216 (blue squares)

Enlarge
  • Infliximab Antibody | AbD19376 thumbnail image 1
  • Infliximab Antibody | AbD19376 thumbnail image 2
  • Infliximab Antibody | AbD19376 thumbnail image 3
  • Infliximab Antibody | AbD19376 thumbnail image 4
  • Infliximab Antibody | AbD19376 thumbnail image 5
  • Infliximab Antibody | AbD19376 thumbnail image 6
  • Human anti Infliximab
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-infliximab antibody is a non-neutralizing anti-idiotypic monovalent Fab antibody; it is ideal for bioanalytical assays for infliximab and biosimilars. It does not inhibit the binding of infliximab to TNF alpha and so detects total drug. It is recommended as PK bridging ELISA capture antibody.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD19376
    • Isotype
      HuCAL Fab monovalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA214Edatasheet pdfdatasheet pdf0.1 mg
      HCA214
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
      -
      • Human anti Infliximab, clone AbD19376, is an anti-idiotypic antibody that specifically recognizes the infliximab monoclonal antibody and does not inhibit the binding of infliximab to its target, Tumor Necrosis Factor Alpha (TNFα). It can be used in bioanalytical assays to measure the levels of infliximab and biosimilar products in patient samples.

        Clone AbD19376 recognizes both the free infliximab monoclonal antibody and infliximab when bound to TNFα (the drug/target complex); it is not paratope-specific and it is non-inhibitory. This antibody can be used as a capture antibody in a pharmacokinetic (PK) assay in the bridging format to detect total drug (free, partially bound and fully bound) with HCA216P as the detection antibody.

        Infliximab (branded as Remicade®) is a chimeric monoclonal antibody drug (IgG1/kappa) that has been approved for treatment of psoriasis, Crohn's disease, ankylosing spondylitis, psoriatic arthritis, rheumatoid arthritis, and ulcerative colitis. It acts by blocking the binding of TNFα to its receptors. Infliximab also induces apoptosis of TNFα expressing T-lymphocytes.

        View a summary of all anti-infliximab antibodies.
      • 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.
      • Reconstitution
      • Preparation
        Metal chelate affinity chromatography
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Infliximab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD=0.7 nM by real time, label-free molecular interaction analysis on immobilized infliximab.
      • 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.
        Remicade is a registered trademark of Janssen Biotech Inc.
      • 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 an indirect ELISA or as a capture antibody in an Infliximab bridging ELISA together HRP-conjugated HCA213 or HCA216 as the detection reagent.
        Protocol: PK bridging ELISA to measure total drug
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Infliximab Antibody Formats

      Formats Clone Applications Sizes available
      Infliximab Antibody : Purified AbD19376 E 0.1 mg
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Rabbit anti DYKDDDDK Tag:HRPAHP1074P0.1 mgE, WB
        AHP1074P
        Rat anti DYKDDDDK Tag:HRPMCA4764P0.1 mgE, P, WB
        MCA4764P
        Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
        MCA5995P

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Mouse anti Histidine Tag:Alk.Phos.MCA1396A0.1 mgE, WB
            MCA1396A
            Goat anti Histidine Tag:BiotinAHP1656B0.1 mgE, WB
            AHP1656B
            Mouse anti Histidine Tag:BiotinMCA1396B0.1 mgE, WB
            MCA1396B
            Mouse anti Human IgG (Fc) CH2 Domain:HRPMCA647P0.2 mgC *, E
            MCA647P
            Human anti InfliximabHCA2130.1 mgE
            HCA213
            Human anti InfliximabHCA2150.1 mgE
            HCA215
            Human anti InfliximabHCA2160.1 mgE
            HCA216
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A
            Recombinant Human TNF AlphaPHP05150 µgE, FN, WB
            PHP051

            Recommended Positive Controls

              Histology Controls

                Write your review

                You may also be interested in...

                You May Also Be Interested In...