Multienzyme detection and in-situ monitoring of enzyme activity by bending CE using quantum dots-based polypeptide substrate

Electrophoresis. 2020 Jun;41(12):1103-1108. doi: 10.1002/elps.202000023. Epub 2020 Mar 3.

Abstract

Multienzyme detection and monitoring enzyme activity in situ are significant for the disease to diagnose. This study aims to develop a quantum dots (QDs)-based nanoprobe Cyanine5-DDDLEVLFQFPGLVPRGSGGHHHHHH-QDs (Cy5-LEVLVP-QD), which is able to detect two enzymes inside a bent capillary using CE. Cy5-LEVLVP and QDs were allowed to bind with each other through metal affinity interaction and then injected the Cy5-LEVLVP-QD complex into a capillary with different bends, followed by related enzyme that can cleave the Cy5-LEVLVP peptide. The fluorescence of Cy5 was excited by QDs due to Förster resonance energy transfer. By monitoring the peaks produced by the original Cy5-LEVLVP-QD complex and a significant fluorescence change, sensitive analysis of two different enzymes was conducted. Therefore, the novel approach of using capillaries with semicircular bends could prove particularly useful for enzyme investigating in disease.

Keywords: CE; Förster resonance energy transfer; enzyme; peptide; quantum dots.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbocyanines / analysis
  • Carbocyanines / chemistry
  • Electrophoresis, Capillary / methods*
  • Enzyme Assays
  • Enzymes* / analysis
  • Enzymes* / metabolism
  • Fluorescence Resonance Energy Transfer / methods*
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / chemistry
  • Peptides / analysis
  • Peptides / chemistry
  • Peptides / metabolism
  • Quantum Dots* / analysis
  • Quantum Dots* / chemistry

Substances

  • Carbocyanines
  • Enzymes
  • Fluorescent Dyes
  • Peptides
  • cyanine dye 5