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Following hybridization on sensor/array platforms by using SPR, elipsometer and MALDI-MS.

Ömür Çelikbıçak1, Kadriye Özlem Hamaloğlu2, Bekir Salih1

  • 1Department of Chemistry, Hacettepe University, Ankara, Turkey.

Nucleosides, Nucleotides & Nucleic Acids
|May 14, 2020
PubMed
Summary

This study introduces a novel method combining Surface Plasmon Resonance (SPR), ellipsometry, and MALDI-Mass Spectrometry (MALDI-MS) for detecting single-strand oligodeoxynucleotides (ssODNs). The integrated approach enables sensitive and specific ssODN detection on functionalized surfaces.

Keywords:
Hybridization sensors/arraysellipsometermatrix-assisted laser desorption/ıonization-mass spectrometrymicro-contact printingself-assemblingsurface plasmon resonance

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Nanotechnology

Background:

  • Accurate detection of single-strand oligodeoxynucleotides (ssODNs) is crucial for various biological and diagnostic applications.
  • Existing methods often face challenges in sensitivity, specificity, or require complex sample preparation.
  • Developing integrated platforms for ssODN detection can enhance analytical capabilities.

Purpose of the Study:

  • To develop and validate a multiplexed methodology for ssODN detection.
  • To integrate Surface Plasmon Resonance (SPR), Ellipsometry (EM), and Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry (MALDI-MS) for enhanced detection.
  • To optimize surface functionalization for controlled probe immobilization and minimize non-specific interactions.

Main Methods:

  • Micro-contact printing (µCP) of probe-ssODNs onto gold-coated surfaces using silicone stamps.
  • Co-immobilization of a modulator-spacer molecule (6-mercapto-1-hexanol) to control probe density and orientation.
  • Real-time monitoring of hybridization using SPR, surface imaging with ellipsometry (IEM), and final detection via MALDI-MS after dehybridization.

Main Results:

  • Successful homogeneous immobilization of probe-ssODNs achieved via µCP.
  • SPR demonstrated successful hybridization monitoring with complete hybridization in 100 minutes.
  • A linear relationship was observed between SPR signal change and target concentration below 1 µm.
  • IEM provided complementary surface imaging data, and MALDI-MS confirmed the detection of hybridized target-ssODNs.

Conclusions:

  • The integrated SPR, EM, and MALDI-MS platform offers a robust and sensitive method for ssODN detection.
  • The developed methodology allows for quantitative analysis and confirmation of target hybridization.
  • This approach holds promise for applications requiring high-throughput and specific nucleic acid detection.