Analytical Sciences

Abstract − Analytical Sciences, 28(11), 1059 (2012).

A Pivot-Hinge-Style DNA Immobilization Method with Adaptable Surface Concentration Based on Oligodeoxynucleotide-Phosphorothioate Chemisorption on Gold Surfaces
Hisao YOSHINAGA,* Koji NAKANO,* Nobuaki SOH,** Ryoichi ISHIMATSU,* and Toshihiko IMATO*
*Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi, Fukuoka 819-0395, Japan
**Department of Agriculture, Saga University, 1 Honjo, Saga 840-8502, Japan
The chemisorption of oligodeoxynucleotide phosphorothioate (s-oligo) is reported. A series of s-oligo DNAs was designed for use as capture probe DNA molecules. The s-oligo DNAs consist of the K-ras gene (5′-GGA GCT GGT GGC-3′) and a dodecamer deoxyriboadenosine, both of which lie on either side of an s-oligo DNA sequence. By primarily focusing on the capture probe DNA having five-successive s-oligo sequences, e37, the immobilization chemistry of e37 was examined; atomic force microscopy achieved the direct visualization of individual molecules on Au(111) substrates, while a series of surface analyses, including IR, ellipsometry, and microgravimetry, showed that the s-oligo functional groups played a pivotal role in the surface-adlayer through the gold-thiol interaction. Interestingly, the amount of immobilization showed a definite relationship with the number of s-oligo linkages introduced, which should be important to regulate the concentration of the capture probe DNA molecules on the surface. Some preliminary studies using ferrocene-modified complementary sequences indicated that electrochemical labeling and readouts were possible.