Huang Tom T, Geng Tao, Akin Demir, Chang Woo-Jin, Sturgis Jennifer, Bashir Rashid, Bhunia Arun K, Robinson J Paul, Ladisch Michael R
School of Chemical Engineering and Laboratory of Renewable Resources Engineering, Purdue University, West Lafayette, Indiana 47907, USA.
Biotechnol Bioeng. 2003 Aug 20;83(4):416-27. doi: 10.1002/bit.10680.
This work describes a simple approach to immobilize functionalized colloidal microstructures onto a C(18)-coated SiO(2) substrate via specific or non-specific bio-mediated interactions. Biotinylated bovine serum albumin pre-adsorbed onto a C(18) surface was used to mediate the surface assembly of streptavidin-coated microbeads (2.8 microm), while a bare C(18) surface was used to immobilize anti-Listeria antibody-coated microbeads (2.8 microm) through hydrophobic interactions. For a C(18) surface pre-adsorbed with bovine serum albumin, hydrophobic polystyrene microbeads (0.8 microm) and positively charged dimethylamino microbeads (0.8 microm) were allowed to self-assemble onto the surface. A monolayer with high surface coverage was observed for both polystyrene and dimethylamino microbeads. The adsorption characteristics of Escherichia coli and Listeria monocytogenes on these microbead-based surfaces were studied using fluorescence microscopy. Both streptavidin microbeads pre-adsorbed with biotinylated anti-Listeria antibody and anti-Listeria antibody-coated microbeads showed specific capture of L. monocytogenes, while polystyrene and dimethylamino microbeads captured both E. coli and L. monocytogenes non-specifically. The preparation of microbead-based surfaces for the construction of microfluidic devices for separation, detection, or analysis of specific biological species is discussed.
这项工作描述了一种通过特异性或非特异性生物介导的相互作用,将功能化胶体微结构固定在C(18)涂层的SiO(2)基板上的简单方法。预先吸附在C(18)表面的生物素化牛血清白蛋白用于介导链霉亲和素包被的微珠(2.8微米)的表面组装,而裸露的C(18)表面则通过疏水相互作用固定抗李斯特菌抗体包被的微珠(2.8微米)。对于预先吸附了牛血清白蛋白的C(18)表面,允许疏水聚苯乙烯微珠(0.8微米)和带正电荷的二甲基氨基微珠(0.8微米)在表面自组装。观察到聚苯乙烯和二甲基氨基微珠都形成了具有高表面覆盖率的单层。使用荧光显微镜研究了大肠杆菌和单核细胞增生李斯特菌在这些基于微珠的表面上的吸附特性。预先吸附了生物素化抗李斯特菌抗体的链霉亲和素微珠和抗李斯特菌抗体包被的微珠都显示出对单核细胞增生李斯特菌的特异性捕获,而聚苯乙烯和二甲基氨基微珠则非特异性地捕获大肠杆菌和单核细胞增生李斯特菌。本文还讨论了用于构建用于分离、检测或分析特定生物物种的微流控装置的基于微珠的表面的制备方法。