Locascio Laurie E, Hong Jennifer S, Gaitan Michael
Analytical Chemistry Division, National Institute of Standards and Technology, Gaithersburg, MD 20899-8394, USA.
Electrophoresis. 2002 Mar;23(5):799-804. doi: 10.1002/1522-2683(200203)23:5<799::AID-ELPS799>3.0.CO;2-P.
Liposomes with encapsulated carboxyfluorescein were used in an affinity-based assay to provide signal amplification for small-volume fluorescence measurements. Microfluidic channels were fabricated by imprinting in a plastic substrate material, poly(ethylene terephthalate glycol) (PETG), using a silicon template imprinting tool. Streptavidin was linked to the surface through biotinylated-protein for effective immobilization with minimal nonspecific adsorption of the liposome reagent. Lipids derivatized with biotin were incorporated into the liposome membrane to make the liposomes reactive for affinity assays. Specific binding of the liposomes to microchannel walls, dependence of binding on incubation time, and nonspecific adsorption of the liposome reagent were evaluated. The results of a competitive assay employing liposomes in the microchannels are presented.
将包裹有羧基荧光素的脂质体用于基于亲和力的分析中，以实现小体积荧光测量的信号放大。使用硅模板压印工具，通过在塑料基材聚对苯二甲酸乙二醇酯（PETG）中压印来制造微流控通道。链霉亲和素通过生物素化蛋白连接到表面，以实现有效固定，同时使脂质体试剂的非特异性吸附最小化。将用生物素衍生化的脂质掺入脂质体膜中，使脂质体对亲和力分析具有反应性。评估了脂质体与微通道壁的特异性结合、结合对孵育时间的依赖性以及脂质体试剂的非特异性吸附。展示了在微通道中使用脂质体进行竞争性分析的结果。
