Poorly soluble peptides can mimic authentic ELISPOT responses

R. Karl Karlsson, Wim Jennes, Kimberly Page-Shafer, Douglas F. Nixon, Barbara Shacklett

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

The ELISPOT assay is a specific, sensitive, quantitative assay for assessing cell-mediated immune responses to a variety of antigens including HIV-1 peptides. In an interferon (IFN)-γ-ELISPOT assay, peripheral blood mononuclear cells (PBMC) from two HIV-1 exposed seronegative (ESN) individuals appeared to respond strongly to an HIV Gag peptide. Analysis of this peptide revealed that it was incompletely dissolved and induced non-specific spot formation, even in the absence of cells. In subsequent experiments, the peptide was found to interact with avidin and the ELISPOT membrane. Filtering the peptide prevented non-specific spot formation. These findings underscore the need for appropriate controls and proper peptide preparation in order to reduce the risk of false-positive ELISPOT responses.

Original languageEnglish (US)
Pages (from-to)89-92
Number of pages4
JournalJournal of Immunological Methods
Volume285
Issue number1
DOIs
StatePublished - Feb 1 2004

Keywords

  • ELISPOT
  • False-positive
  • HIV-1
  • Non-specific
  • Peptide

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

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