Luminescence stability improvement in liposome-based homogeneous luminescence resonance energy transfer

Anal Biochem. 2013 Sep 1;440(1):6-8. doi: 10.1016/j.ab.2013.05.004. Epub 2013 May 20.

Abstract

A stable liposome-based time-resolved luminescence resonance energy transfer (TR-LRET) assay was developed based on the interaction of biotinylated lipids and streptavidin. Eu(3+) ion chelated to 4,4,4-trifluoro-1-(2-naphthalenyl)-1,3-butanedione and trioctylphosphine oxide was incorporated into liposomes. Acceptor-labeled streptavidin bound to biotinylated lipids of the liposomes enables TR-LRET. A stable assay performance was achieved by optimization. High Eu(3+) signal and stability, low variation, and sensitivity below 100 pM for free biotin was achieved by incorporating the chelate into liposomes containing cholesterol in a carbonate buffer. Potentially, the stable assay compared with the assay without cholesterol offers an improved platform to liposome-based detection systems.

Keywords: Biotin; Cholesterol; Energy transfer; Liposome; Streptavidin; Time-resolved luminescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin / analysis
  • Biotin / chemistry*
  • Cholesterol / analysis
  • Cholesterol / chemistry*
  • Europium / analysis
  • Europium / chemistry*
  • Fluorescence Resonance Energy Transfer / methods*
  • Ketones / analysis
  • Ketones / chemistry*
  • Liposomes / analysis
  • Liposomes / chemistry*
  • Luminescence*
  • Naphthalenes / analysis
  • Naphthalenes / chemistry*
  • Streptavidin / analysis
  • Streptavidin / chemistry*

Substances

  • 4,4,4-trifluoro-1-(2-naphthalenyl)-1,3-butanedione
  • Ketones
  • Liposomes
  • Naphthalenes
  • Europium
  • Biotin
  • Streptavidin
  • Cholesterol