Characterization of molecular interactions of an immobilized biotinylated monolayer and streptavidin-coated microspheres by bond-rupture scanning.
Bond-rupture approach has been used in the understanding of biomolecular interactions of highly specific recognition, e.g., an antibody and its antigen, by a functionalized and self-assembled monolayer (SAM). One of the most challenging issues of diagnostics is to distinguish between true binding and the ever-present non-specific binding in which a species gives false results in conventional affinity methods. In this study, bond-rupture scanning was proposed to characterize bindings by introducing energy mechanically through displacement of a resonant quartz crystal. This system was able to measure the resonant frequency difference, due to mass changes and bond breakages between supramolecular interaction of biotinylated SAM and streptavidin-coated polystyrene microsphere (SCPM). Both 2-μm and 4-μm of SCPMs revealed two recognized desorption patterns at 4 V and 2 V amplitudes respectively. It rapidly provided confirmation of the presence of a target analyte. From this study, it can be shown that an established approach of dynamic bond-rupture scanning can be adopted as a promising diagnostic tool for investigating various interactions of bacteria or virus on an immobilized biomolecular surface by measuring the characteristic level of mechanical energy required to break bonds.
School of Materials Science & Engineering, Southwest Jiaotong University, Chengdu 610031, Sichuan, PR China. yongyuan@swjtu.edu.cn
Yong J
YJ
Yuan
Yudi
Y
Zhao
Wei L
WL
Xu
D019921:Biotinylation; D004800:Enzymes, Immobilized / Q000737:chemistry*; D008863:Microspheres; D008954:Models, Biological; D058525:Quartz Crystal Microbalance Techniques; D019809:Streptavidin / Q000737:chemistry*
D004800:Enzymes, Immobilized; D019809:Streptavidin