Synthetic peptide-based elisa and elispot assay for identifying autoantibody epitopes

Judit Pozsgay, Eszter Szarka, Krisztina Huber, Fruzsina Babos, Anna Magyar, Ferenc Hudecz, Gabriella Sarmay

Research output: Chapter in Book/Report/Conference proceedingChapter

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-specifi c autoreactive B cells in a pre-stimulated culture of patients’ lymphocytes.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages223-233
Number of pages11
DOIs
Publication statusPublished - Jan 1 2016

Publication series

NameMethods in Molecular Biology
Volume1352
ISSN (Print)1064-3745

    Fingerprint

Keywords

  • Anti-citrullinated peptide antibodies
  • B cell
  • Citrulline-peptide
  • Diagnosis
  • ELISA
  • ELISpot
  • Multipin ELISA
  • Rheumatoid arthritis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Pozsgay, J., Szarka, E., Huber, K., Babos, F., Magyar, A., Hudecz, F., & Sarmay, G. (2016). Synthetic peptide-based elisa and elispot assay for identifying autoantibody epitopes. In Methods in Molecular Biology (pp. 223-233). (Methods in Molecular Biology; Vol. 1352). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3037-1_17