A mixed-phase immunoassay based on simultaneous binding of fluorescently tagged and PNA-conjugated peptide epitopes on antibodies: quantification on PNA-coated microparticles

Bioconjug Chem. 2006 Jul-Aug;17(4):1063-8. doi: 10.1021/bc0600315.

Abstract

A mixed-phase immunoassay based on simultaneous binding of an antibody to its fluorescently tagged peptide epitope and a PNA conjugate of the same peptide has been developed. As a fluorescent marker, a europium(III) chelate allowing time-resolved measurement from a single particle has been employed. The ternary complex formed in solution is immobilized by Watson-Crick base-pairing to a microparticle bearing a PNA sequence complementary to that present in the complex. The concentration of the antibody in the sample may then be determined by a single particle measurement. Accordingly, different antibodies may in principle be addressed by sequence-specific hybridization to different categorized microparticles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antibodies / chemistry
  • Antibodies / metabolism*
  • Epitopes / metabolism*
  • Fluorescent Dyes / metabolism*
  • Immunoassay / methods*
  • Molecular Sequence Data
  • Peptide Nucleic Acids / metabolism*

Substances

  • Antibodies
  • Epitopes
  • Fluorescent Dyes
  • Peptide Nucleic Acids