Characterization of PNA and DNA Immobilization and Subsequent Hybridization with DNA Using Acoustic-Shear-Wave Attenuation Measurements

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Characterization of PNA and DNA Immobilization and Subsequent Hybridization with DNA Using Acoustic-Shear-Wave Attenuation Measurements

Höök, Fredrik, 1966 (author): Chalmers tekniska högskola,Chalmers University of Technology

Ray, Arghya (author): Chalmers tekniska högskola,Chalmers University of Technology

Nordén, Bengt, 1945 (author): Chalmers tekniska högskola,Chalmers University of Technology

Kasemo, Bengt Herbert, 1942 (author): Chalmers tekniska högskola,Chalmers University of Technology

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2001-12-01
2001
English.
In: Langmuir. - : American Chemical Society (ACS). - 1520-5827 .- 0743-7463. ; 17:26, s. 8305-8312

Related links:: http://dx.doi.org/10...; show more...; https://research.cha...; https://doi.org/10.1...; show less...

Journal article (peer-reviewed)

Abstract Subject headings

We report here how the quartz crystal microbalance with dissipation monitoring (QCM-D) technique, simultaneously measuring changes in the induced energy dissipation, D (cf. viscoelastic properties), and the frequency, f (cf. coupled mass), can be used to characterize the bound state of single-stranded peptide nucleic acid (PNA) and deoxyribose nucleic acid (DNA) in relation to their ability to function as selective probe(s) for fully complementary and single-mismatch DNA. The possibility to use the QCM-D technique for detection of binding kinetics and structural differences in the formed duplexes is also presented. We found that thiol-PNA and thiol-DNA attached via a sulfur group directly on a bare-gold surface are less efficient as probes for DNA than are biotin-PNA and biotin-DNA, coupled on top of a two-dimensional (2-D) arrangement of streptavidin, formed on a biotinylated phospholipid bilayer on a SiO2 surface. The fully complementary and singly mismatched DNA oligomers hybridize with the immobilized PNA and DNA. A single mismatch is discriminated via a significant difference in the binding and dissociation kinetics, demonstrating a high selectivity and thus successful immobilization of functional single strands. The observed ratios between hybridization-induced energy dissipation (DeltaD) and the frequency shift (Deltaf) made it possible to discriminate thiol-PNA directly attached to a gold surface from biotin-PNA coupled to the streptavidin 2-D arrangement, where the former were shown to be inefficient for detecting subsequent hybridization. Structural differences of the immobilized layers composed of biotin-PNA-DNA and biotin-DNA-DNA were clearly reflected by the DeltaD and Deltaf response.

Subject headings

NATURVETENSKAP -- Kemi -- Fysikalisk kemi (hsv//swe)
NATURAL SCIENCES -- Chemical Sciences -- Physical Chemistry (hsv//eng)

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NATURAL SCIENCES: NATURAL SCIENCES; and Chemical Science ...; and Physical Chemist ...

Articles in the publication: Langmuir

By the university: Chalmers University of Technology

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