New immobilisation method for oligonucleotides on electrodes enables highly-sensitive, electrochemical label-free gene sensing

Biosens Bioelectron. 2017 Nov 15:97:128-135. doi: 10.1016/j.bios.2017.05.049. Epub 2017 Jun 1.

Abstract

We present a versatile and facile procedure for the immobilisation of bioprobe molecules to an electrochemical sensing element. We eliminate lengthy preparation procedures for direct functionalisation of electrode surfaces by pre-attaching probe molecules to carboxylic acid bearing termonomers of pyrrole phenylenes or thiophene phenylenes. We demonstrate that these conjugates can be electrodeposited at low potentials to form nano-scale porous, electroactive conducting polymer films, exposing the bioprobe and retaining activity and specificity for binding, exemplified here with DNA sensors. The electrochemical reaction impedance for Fe(CN)63-/4- on oligonucleotide-modified electrodes showed remarkable (down to aM) detection sensitivity for target DNA sequences present in solution. Cross-sensitivity to non-complementary target sequences is small and multi-target arrays are easily made. There is no need for labelling of either probe or target oligonucleotide.

Keywords: Biomolecule immobilisation; Conducting polymers; Electrochemical biosensors; Label-free sensing.

MeSH terms

  • Biosensing Techniques / methods*
  • Carboxylic Acids / chemistry
  • DNA / analysis*
  • Electrochemical Techniques / methods*
  • Electrodes
  • Electroplating / methods
  • Oligonucleotide Probes / chemistry*
  • Pyrroles / chemistry*
  • Thiophenes / chemistry*

Substances

  • Carboxylic Acids
  • Oligonucleotide Probes
  • Pyrroles
  • Thiophenes
  • DNA