Amperometric Detection of Apoptosis by using p-Methoxyaniline-Conjugated Substrate for Caspase-3

Sun Sixiang, Kumi Y. Inoue, Shusaku Shiomoto, Shinichiro Takano, Kosuke Ino, Hitoshi Shiku, Tomokazu Matsue

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

This manuscript reports on a novel substrate for the amperometric detection of cellular apoptosis, Asp-Glu-Val-Asp-p-methoxyaniline (DEVD-pMA). This substrate showed electrochemical inertia from −0.2 to 0.7 V versus Ag/AgCl in phosphate-buffered saline (PBS), but was cleaved by caspase-3, releasing pMA, which was detected with cyclic voltammetry and chronoamperometry. Compared with p-aminophenol (pAP), pMA showed lower toxicity to the human hepatocellular carcinoma cell line (HepG2) and higher stability in PBS. By using this substrate, we successfully detected caspase-3 in the range of 0.31–2.5×10−2 units/mL and cellular apoptosis in HepG2 cells by using simple amperometry without cell lysis. This work will help to develop a simple method for the electrochemical monitoring of apoptosis, which can be applied in biological research and drug development.

Original languageEnglish
Pages (from-to)941-946
Number of pages6
JournalChemElectroChem
Volume4
Issue number4
DOIs
Publication statusPublished - 2017 Apr 1

Keywords

  • amperometric detection
  • apoptosis
  • biosensing
  • caspase-3
  • peptide substrate

ASJC Scopus subject areas

  • Catalysis
  • Electrochemistry

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