Poorly soluble peptides can mimic authentic ELISPOT responses. Academic Article uri icon

Overview

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)-gamma-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.

publication date

  • February 1, 2004

Research

keywords

  • Enzyme-Linked Immunosorbent Assay
  • Peptides

Identity

Scopus Document Identifier

  • 0442292284

Digital Object Identifier (DOI)

  • 10.1016/j.jim.2003.11.013

PubMed ID

  • 14871537

Additional Document Info

volume

  • 285

issue

  • 1