School of Pharmacy and Department of Chemistry, Fudan University, Shanghai, P R China.
Electrophoresis. 2010 Sep;31(18):3070-3. doi: 10.1002/elps.201000108.
In this report, an inflation bulb-driven microfluidic reactor was developed for IR-accelerated proteolysis. This novel proteolysis system mainly consisted of an inflation bulb-driving system, a simple cross-PMMA microchip, and a temperature-controllable IR radiation system. The gas pressure generated from an inflation bulb was employed to drive protein and trypsin solutions to flow into the main channel of the microchip via two capillaries and the injection channel. When the two solutions were mixed in the channel, the protein was rapidly digested by trypsin under IR radiations. The peptides in the digests accumulated in the product reservoir of the microchip were subsequently identified by MS. The feasibility and performance of this unique system were demonstrated by digesting hemoglobin and lysozyme. The results indicated that IR radiation could significantly enhance the on-chip proteolysis and the digestion time was substantially reduced to 5 min. The present proteolysis setup is simple and efficient and will find wide applications in high-throughput protein digestion.
在本报告中,开发了一种基于膨胀球驱动的微流控反应器,用于 IR 加速蛋白水解。这个新颖的蛋白水解系统主要由膨胀球驱动系统、简单的 PMMA 十字微芯片和温度可控的 IR 辐射系统组成。膨胀球产生的气压用于将蛋白质和胰蛋白酶溶液通过两个毛细管和注入通道驱动到微芯片的主通道中。当两种溶液在通道中混合时,蛋白质在 IR 辐射下被胰蛋白酶迅速水解。微芯片产品储存器中的消化产物中的肽随后通过 MS 进行鉴定。通过消化血红蛋白和溶菌酶验证了该独特系统的可行性和性能。结果表明,IR 辐射可以显著增强芯片上的蛋白水解作用,并且消化时间大大缩短至 5 分钟。目前的蛋白水解装置简单高效,将在高通量蛋白消化中得到广泛应用。