Lee Jeong Hoon, Han Jongyoon
Department of Electrical Engineering, Kwangwoon University, 447-1, Wolgye, Nowon, Seoul, South Korea 139-701.
Microfluid Nanofluidics. 2010 Oct 1;9(4):973-979. doi: 10.1007/s10404-010-0598-z.
Here, we report a new method of concentration-enhanced binding kinetics for a rapid immunoassay screening test on a gold surface in a poly(dimethylsiloxane) (PDMS) microfluidic chip format. The use of alkylthiolate self-assembled monolayers on gold surfaces of a PDMS-glass microchip resulted in accelerated binding kinetics of Human chorionic gonadotropin (hCG) at an electrokinetic trapping zone. We used a PBS solution (buffer concentration ~ 150 mM), not a dibasic buffer system (~10 mM), for the dynamic preconcentrating operation and the preconcentration of cy3 labeled streptavidin onto biotinylated Au surface revealed that the binding kinetics of the protein were linearly proportional to the concentration profile of the preconcentration plug. We showed rapid detection of hCG in the clinical range with a shorten assay time of 10 min. Also, we demonstrated that the amount of sample needed were detection was decreased from ~4 mL to ~25 μL in the standard serum tests. The enhanced binding kinetics between hcG Ag-Ab via preconcentration showed good feasibility for use in a rapid immunoassay screening test.
在此，我们报道了一种用于在聚二甲基硅氧烷（PDMS）微流控芯片形式的金表面上进行快速免疫分析筛选测试的浓度增强结合动力学新方法。在PDMS-玻璃微芯片的金表面上使用硫醇盐自组装单分子层，导致人绒毛膜促性腺激素（hCG）在电动捕获区的结合动力学加快。我们使用PBS溶液（缓冲液浓度约150 mM）而非二元缓冲系统（约10 mM）进行动态预浓缩操作，并且将cy3标记的链霉亲和素预浓缩到生物素化的金表面上，结果表明蛋白质的结合动力学与预浓缩塞的浓度分布呈线性比例关系。我们展示了在临床范围内对hCG的快速检测，检测时间缩短至10分钟。此外，我们证明在标准血清测试中，检测所需的样品量从约4 mL减少至约25 μL。通过预浓缩增强的hCG抗原-抗体之间的结合动力学在快速免疫分析筛选测试中显示出良好的可行性。
