Sol-gel-derived Poly（dimethylsiloxane） Enzymatic Reactor for Microfluidic Peptide Mapping

作     者：吴会灵 杨芃原 范国荣 田玉平 陆豪杰 金红 

作者机构：School of Pharmacy Second Military Medical University the People's Liberation Army Shanghai 200433 China Department of Chemistry Research Center for Proteome Fudan University Shanghai 200433 China 

出 版 物：《Chinese Journal of Chemistry》 (中国化学（英文版）)

年 卷 期：2006年第24卷第7期

页      面：903-909页

核心收录：

学科分类：0710[理学-生物学] 071010[理学-生物化学与分子生物学] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 0703[理学-化学] 

基　　金：Project supported by the National Natural Science Foundation of China (Nos. 30572227  20299030)  Shanghai Science Research Foundation (No 03JC14005)  National Basic Research Priority Program (No. 001 CB510202) and National High Technology Key Project (No. 2002BAC11A11) 

主　　题：microfluidic enzymatic reactor silica sol-gel matrix-assisted laser desorption ionization time-of-flight mass spectrometry electrospray ionization mass spectroscopy high-throughput protein identification 

摘      要：The silica-based poly（dimethylsiloxane） （PDMS） microfluidic enzymatic reactor was reported along with its analytical features in coupling with MALDI TOF and ESI MS. Microfluidic chip was fabricated using PDMS casting and O2-plasma techniques, and used for the preparation of enzymatic reactor. Plasma oxidation for PDMS enabled the channel wall of microfluidics to present a layer of silanol （SiOH） groups. These SiOH groups as anchors onto the microchannel wall were linked covalently with the hydroxy groups of trypsin-encapsulated sol matrix. As a result, the leakage of sol-gel matrix from the microchannel was effectively prevented. On-line protein analysis was performed with the microfluidic enzymatic reactor by attachment of stainless steel tubing electrode and replaceable tip, The success of trypsin encapsulation was investigated by capillary electrophoresis （CE） detection, and MALDI TOF and ESI MS analysis. The lab-made device provided excellent extent of digestion even at the fast flow rate of 7.0 μL/min with very short residence time of ca. 2 s. In addition, the encapsulated trypsin exhibits increased stability even after continuous use. These features are the most requisite for high-throughput protein identification.