A europium fluoroimmunoassay for measuring peptide binding to MHC class I molecules

Peter E. Jensen, Joseph C. Moore, Aron E. Lukacher

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

A fluoroimmunoassay employing europium-streptavidin and time-resolved fluorimetry was used to measure binding of biotin-labeled peptides to H- 2D(k) molecules. A fluorescein-labeled octameric peptide from the middle T (MT) protein of mouse polyoma virus was identified that specifically binds to purified D(k) using a previously-established assay based on size exclusion chromatography. A europium immunoassay was adapted to measure binding of a biotin-derivative of this peptide to purified D(k). The assay was found to be sensitive and highly specific. Binding was optimal at pH 5.0 and 24-27°C, and it was not enhanced in the presence of additional β2-microglobulin (β2m). Excellent results were also obtained in experiments with fixed cells that express D(k). This assay is expected to be useful for high volume, routine analysis of peptide binding to MHC class I molecules.

Original languageEnglish (US)
Pages (from-to)71-80
Number of pages10
JournalJournal of Immunological Methods
Volume215
Issue number1-2
DOIs
StatePublished - Jun 1 1998

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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