Nucleic acid-controlled quantum dots aggregation: A label-free fluorescence turn-on strategy for alkaline phosphatase detection

Talanta. 2017 Jul 1:169:64-69. doi: 10.1016/j.talanta.2017.03.063. Epub 2017 Mar 22.

Abstract

Based on the controlled aggregation of quantum dots (QDs), a valid, reliable, and label-free fluorescence turn-on strategy is established for the detection of alkaline phosphatase activity. The aqueous solution of anionic QDs exhibits intense fluorescence. However, the addition of cationic polymer (poly-1) significantly quenched the fluorescence of the QDs via their induced aggregation. While short 3'-phosphorylated DNA (DNA-P) could not be extended by terminal deoxynucleotidyl transferase (TdT) and therefore, fluorescence of the QDs was recovered negligibly. The effective elimination of phosphate group of DNA-P in the presence of alkaline phosphatase (ALP) produced 3'-OH termini and the resulting DNA could be sufficiently extended by TdT. The presence of greater binding strength between the elongated DNA and poly-1 is very crucial to compete with the poly-1/QDs aggregates and release the QDs. Turned-on fluorescence emission is observed due to the efficient release of the QDs. A novel strategy for alkaline phosphatase detection is therefore established. Our method is quite sensitive and selective, as low as 0.1 mU/mL ALP can be easily detected.

Keywords: Alkaline phosphatase; Controlled-aggregation; Fluorescence; Nucleic acid; Quantum dots; Terminal deoxynucleotidyl transferase.

MeSH terms

  • Alkaline Phosphatase / analysis*
  • Fluorescence*
  • Fluorescent Dyes / chemistry*
  • Humans
  • Nucleic Acids / chemistry*
  • Quantum Dots*
  • Spectrometry, Fluorescence

Substances

  • Fluorescent Dyes
  • Nucleic Acids
  • Alkaline Phosphatase