Characterization of DNA immobilization and subsequent hybridization on a 2D arrangement of streptavidin on a biotin-modified lipid bilayer supported on SiO2

被引:287
作者
Larsson, C [1 ]
Rodahl, M
Höök, F
机构
[1] Chalmers Univ Technol, Dept Appl Phys, S-41296 Gothenburg, Sweden
[2] Univ Gothenburg, S-41296 Gothenburg, Sweden
[3] Q Sense AB, S-42677 Vastra Frolunda, Sweden
关键词
D O I
10.1021/ac034269n
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We show how the water content (and effective density) of thin adsorbed films composed of biomolecules can be determined using combined quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) analysis. In particular, these techniques, combined with theoretical treatment using a Voigt-based viscoelastic model, were applied to analyze the state of surface immobilized single stranded biotin-modified probe DNA (b-DNA) coupled via streptavidin to a biotindoped supported phospholipid bilayer (b-SPB)). From a proper analysis, it is demonstrated how changes in effective thickness, delta(f), and the viscoelastic components (shear viscosity, eta(f), and shear elasticity, mu(f))) can be obtained during both DNA immobilization and hybridization with single stranded fully complementary target DNA. In particular, it is demonstrated how this type of analysis can be used to control the state of streptavidin arrangement for improved measurements of DNA hybridization kinetics. The latter is demonstrated by identifying a surface-coverage dependent viscoelastic behavior of immobilized b-DNA, which is shown to influence the hybridization efficiency.
引用
收藏
页码:5080 / 5087
页数:8
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