Surface plasmon resonance spectroscopy and quartz crystal microbalance study of streptavidin film structure effects on biotinylated DNA assembly and target DNA hybridization.
Surface plasmon resonance (SPR) spectroscopy is employed for the study of biotinylated DNA assembly on streptavidin modified gold surfaces for target DNA hybridization. Two immobilization strategies are involved for constructing streptavidin films, namely, (1) physical adsorption on biotin-containing thiol treated surfaces through biotin-streptavidin links and (2) covalent attachment to 11-mercaptoundecanoic acid (MUA) treated surfaces through amine coupling. To understand the structural properties of the streptavidin films, a quartz crystal microbalance with energy dissipation monitoring (QCM-D) is used to monitor the streptavidin immobilization procedures. The simultaneously measured frequency (Deltaf) and dissipation factor (DeltaD) changes, together with the SPR angle shifts (Deltatheta), suggest that the streptavidin film assembled on the biotin-containing surface is highly rigid with a well-ordered structure while the streptavidin film formed through amine coupling is highly dissipative and less structured. The subsequent biotinylated DNA (biotin-DNA) assembly and target hybridization results show that the streptavidin film structure has distinct effects on the biotin-DNA binding amount. On the streptavidin matrix, not only the probe DNA density but also the strand orientation mediated by the streptavidin films has distinct effects on hybridization efficiency. Particularly, the molecularly ordered streptavidin films formed on the biotin-containing surfaces ensure a well-ordered DNA assembly, which in turn allows for a higher efficiency in target DNA capture and for a higher sensitivity in the hybridization analysis when compared to the biotin-DNA assembled on the less structured streptavidin films formed through amine coupling.
Su
Xiaodi
X
Institute of Materials Research and Engineering, 3 Research Link, Singapore 117602, and Max-Planck-Institut für Polymerforschung, Ackermannweg 10, 55128 Mainz, Germany.
Wu
Ying-Ju
YJ
Robelek
Rudolf
R
Knoll
Wolfgang
W
D001483:Base Sequence; D001710:Biotin / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes; D009693:Nucleic Acid Hybridization; D019809:Streptavidin / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods*
D015342:DNA Probes; D001710:Biotin; D004247:DNA; D019809:Streptavidin