FRET-based homogeneous immunoassay on a nanoparticle-based photonic crystal

Anal Bioanal Chem. 2015 Jul;407(18):5243-7. doi: 10.1007/s00216-015-8708-0. Epub 2015 May 10.

Abstract

The potential of fluorescence resonance energy transfer (FRET) in a photonic crystal (PC) nanostructured array to enhance the speed and sensitivity of a protein-based immunoassay was tested. Forty-nanometer carboxylated particles conjugated with donor-labeled capture antibodies were trapped by electrophoresis and used as a FRET energy donor. The PC array was able to enhance fluorescent excitation and emission by phase matching. To provide a proof of concept for this FRET-based homogeneous assay on a PC chip, an immunoassay was tested with a simple immunoglobulin G (IgG)-based reaction. A standard curve was generated by testing two different antibody reaction times: 20 min. and 1 min. The results were compared directly to those obtained from a FRET assay that used a modern, high-sensitivity plate reader with a 96-well plate and a reaction time of 1 h. The rabbit-IgG detection limits of the FRET-based homogeneous assay on the PC were 0.001 and 0.1 μg/mL for incubation times of 20 and 1 min, respectively; the sensitivities were 10(3) and 10 times better than the 96-well plate reader, respectively. Thus, FRET on a PC immunoplatform was shown to be a facile, effective, rapid, and sensitive detection technology.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Equipment Design
  • Fluorescence Resonance Energy Transfer / economics
  • Fluorescence Resonance Energy Transfer / instrumentation
  • Fluorescence Resonance Energy Transfer / methods*
  • Immunoassay / economics
  • Immunoassay / instrumentation
  • Immunoassay / methods*
  • Immunoglobulin G / analysis*
  • Limit of Detection
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure
  • Photons
  • Rabbits
  • Time Factors

Substances

  • Immunoglobulin G