IUVSTA 15th International Vacuum Congress (IVC-15), AVS 48th International Symposium (AVS-48), 11th International Conference on Solid Surfaces (ICSS-11)
    Biomaterials Monday Sessions
       Session BI-MoM

Paper BI-MoM7
Two-dimensional Arrangements of Streptavidin on Biotin-doped Supported Lipid Bilayars Optimized for DNA-DNA Hybridization

Monday, October 29, 2001, 11:40 am, Room 102

Session: Molecular Recognition
Presenter: F. Höök, Chalmers University of Technology, Sweden
Authors: C. Larsson, Chalmers University of Technology, Sweden
E. Fridell, Chalmers University of Technology, Sweden
B. Kasemo, Chalmers University of Technology, Sweden
F. Höök, Chalmers University of Technology, Sweden
Correspondent: Click to Email
In this work the quartz crystal microbalance with dissipation monitoring (QCM-D), surface plasmon resonance (SPR) and ATR-FTIR techniques were used to follow the 2-D arrangement of streptavidin on a biotinylated phospholipid bilayer supported on a SiO2 surface, for optimized capture of single stranded biotin-DNA (15- and 30-mer) and subsequently hybridization-kinetics measurement of their reaction with mixed-sequences of DNA (15- and 30-mer) with various degrees of mismatch. The QCM-D and ATR-FTIR data suggest that streptavidin rearranges into a more rigid and oriented structure at approximately 50 % coverage, interpreted as onset of crystallization. Interestingly, immobilization of biotin-DNA followed by subsequent hybridization prior to this stage results in up to a two-fold more rapid association kinetics compared with immobilization and hybridization at full coverage of streptavidin. Our results demonstrate how real-time control of the immobilization step can be efficiently used to minimize the influence from lateral interactions for surface-based biorecognition detection, and thus optimize hybridization kinetics measurements. The QCM-D data were also used to quantify variations in the viscoelastic properties of the formed layers of DNA and DNA duplexes, which were further correlated to the obtained results onhybridization kinetics.