A nuclease-assisted label-free aptasensor for fluorescence turn-on detection of ATP based on the in situ formation of copper nanoparticles

Biosens Bioelectron. 2017 Jan 15:87:760-763. doi: 10.1016/j.bios.2016.09.029. Epub 2016 Sep 9.

Abstract

Owing to their promising advantages in biochemical analysis, aptamer-based sensing systems for the fluorescence detection of important biomolecules are being extensively investigated. Herein, we propose a turn-on fluorescent aptasensor for label-free detection of adenosine triphosphate (ATP) by utilizing the in situ formation of copper nanoparticles (CuNPs) and the specific digestion capability of exonuclease I (Exo I). In this assay, the addition of ATP can effectively hinder the digestion of aptamer-derived oligonucleotides due to the G-quadruplex structure. Accordingly, the remaining poly thymine at 5'-terminus of substrate DNA can serve as an efficient template for red-emitting fluorescent CuNPs with a Mega-Stokes shifting in buffered solution, which can be used to evaluate the concentration of ATP. This method is cost-effective and facile, because it avoids the use of traditional dye-labeled DNA strands and complex operation steps. Under optimized conditions, this method achieves a selective response for ATP with a detection limit of 93nM, and exhibits a good detection performance in biological samples.

Keywords: Adenosine triphosphate; Aptamer; Copper nanoparticles; Fluorescence detection; Lable-free; Turn-on.

MeSH terms

  • Adenosine Triphosphate / analysis*
  • Aptamers, Nucleotide / chemistry*
  • Biosensing Techniques / methods*
  • Copper / chemistry*
  • Culture Media / chemistry
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • G-Quadruplexes
  • Limit of Detection
  • Metal Nanoparticles / chemistry*
  • Metal Nanoparticles / ultrastructure
  • Spectrometry, Fluorescence / methods

Substances

  • Aptamers, Nucleotide
  • Culture Media
  • Fluorescent Dyes
  • Copper
  • Adenosine Triphosphate