Biointerphases

Journal for Biophysical Chemistry

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Open Access

Effect of ionic strength on PNA-DNA hybridization on surfaces and in solution

  • Hyeyoung Park1,
  • Andrea Germini2,
  • Stefano Sforza2,
  • Roberto Corradini2,
  • Rosangela Marchelli2 and
  • Wolfgang Knoll1Email author
Biointerphases2:20200080

https://doi.org/10.1116/1.2746871

Received: 29 January 2007

Accepted: 8 May 2007

Abstract

Peptide nucleic acids (PNAs) are mimics of oligonucleotides containing a neutral peptidelike backbone and are able to bind complementary DNA targets with high affinity and selectivity. In order to investigate the effect of the ionic strength of the buffer solution, hybridization experiments with PNAs as (catcher) probes and DNAs as target oligonucleotides were performed in different salt solutions. Surface plasmon field-enhanced fluorescence spectroscopy was employed for real-time monitoring of DNA hybridizations to surface bound PNA. Probes with three different strand lengths were immobilized by self-assembly on the sensor surface. By introducing Cy5-labeled DNA targets the affinity constants, K A=k on (association)/k off (dissociation), were determined for fully complementary (MM0) as well as for single base mismatched (MM1) duplexes. Furthermore, the thermal stability of each duplex was determined by measuring melting curves in solution which was then compared to the kinetic and affinity parameters determined for the surface hybridization reactions. The results indicate that ions do not play a significant role for the PNA/DNA hybridization kinetics at surfaces. However, changes in the configuration of the PNA/DNA duplex due to the ionic strength variations influence the fluorescence yield drastically.