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Synthetic Peptide-Based ELISA and ELISpot Assay for Identifying Autoantibody Epitopes

  • Protocol
Peptide Microarrays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1352))

Abstract

Enzyme-linked immunosorbent assay (ELISA) is an invaluable diagnostic tool to detect serum autoantibody binding to target antigen. To map the autoantigenic epitope(s), overlapping synthetic peptides covering the total sequence of a protein antigen are used. A large set of peptides synthesized on the crown of pins can be tested by Multipin ELISA for fast screening. Next, to validate the results, the candidate epitope peptides are resynthesized by solid-phase synthesis, coupled to ELISA plate directly, or in a biotinylated form, bound to neutravidin-coated surface and the binding of autoantibodies from patients’ sera is tested by indirect ELISA. Further, selected epitope peptides can be applied in enzyme-linked immunospot assay to distinguish individual, citrullinated peptide-specific autoreactive B cells in a pre-stimulated culture of patients’ lymphocytes.

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Author information

Authors and Affiliations

  1. Department of Immunology, Eötvös Loránd University, Pázmány Péter sétány 1/c, Budapest, 1117, Hungary

    Judit Pozsgay, Eszter Szarka, Krisztina Huber & Gabriella Sarmay

  2. Department of Organic Chemistry, Eötvös Loránd University, Budapest, Hungary

    Ferenc Hudecz

  3. MTA-ELTE Research Group of Peptide Chemistry, Hungarian Academy of Sciences, Budapest, Hungary

    Fruzsina Babos, Anna Magyar & Ferenc Hudecz

Authors
  1. Judit Pozsgay
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  2. Eszter Szarka
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  3. Krisztina Huber
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  4. Fruzsina Babos
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  5. Anna Magyar
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  6. Ferenc Hudecz
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  7. Gabriella Sarmay
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Corresponding author

Correspondence to Gabriella Sarmay .

Editor information

Editors and Affiliations

  1. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marina Cretich

  2. Consiglio Nazionale delle Ricerche, Milano, Italy

    Marcella Chiari

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Pozsgay, J. et al. (2016). Synthetic Peptide-Based ELISA and ELISpot Assay for Identifying Autoantibody Epitopes. In: Cretich, M., Chiari, M. (eds) Peptide Microarrays. Methods in Molecular Biology, vol 1352. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3037-1_17

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  • DOI: https://doi.org/10.1007/978-1-4939-3037-1_17

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3036-4

  • Online ISBN: 978-1-4939-3037-1

  • eBook Packages: Springer Protocols

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