Reliable ferrocenyloligonucleotide-immobilized electrodes and their application to electrochemical DNase I assay

Anal Chim Acta. 2009 Jul 10;645(1-2):30-5. doi: 10.1016/j.aca.2009.04.047. Epub 2009 May 6.

Abstract

A ferrocenyloligonucleotide (FcODN) having contiguous cytosine bases was immobilized effectively and reproducibly on a gold electrode furnished with a self-assembled monolayer (SAM) having an N-hydroxysuccinimide-activated carboxylic acid. The resulting electrode was used as a sensor chip in DNase I assay. Thus, the current response of the modified electrode decreased upon addition of DNase I, demonstrating that the phosphodiester bonds of FcODN were cleaved. The DNase I activity assessed by Deltai defined as (i0-i)/i0, where i0 and i represent the current before and after DNase I treatment, respectively, was found to be reproducible with a standard deviation not greater than 9%. The DNase I can be quantitated in the range of 10(-5) to 10(-3) units microL(-1) with a detection limit of 10(-5) units microL(-1) with this sensor chip. The current signal of the FcODN electrode was stable to storage in Biopak water up to 16 days with a 30% signal decrease over this period. DNase I activity in human sera was also determined successfully with this sensor chip.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / economics
  • Biosensing Techniques / methods*
  • Carbodiimides / chemistry*
  • Deoxyribonuclease I / analysis*
  • Deoxyribonuclease I / blood
  • Electrochemical Techniques / economics
  • Electrochemical Techniques / methods*
  • Electrodes
  • Ferrous Compounds / chemistry*
  • Gold / chemistry
  • Humans
  • Oligonucleotides / chemical synthesis
  • Oligonucleotides / chemistry*
  • Reproducibility of Results
  • Time Factors

Substances

  • Carbodiimides
  • Ferrous Compounds
  • Oligonucleotides
  • ferrocenyl carbodiimide
  • Gold
  • Deoxyribonuclease I