整体固定化酶反应器的研制

Construction of a monolithic trypsin enzymatic microreactor

制作了微型整体柱型的固定化酶反应器。在500μm内径毛细管内,以乙烯基三甲氧基硅烷处理形成端基烯键,采用原位合成法,以甲基丙烯酸2-羟乙酯为功能单体,以乙二醇二甲基丙烯酸酯为交联剂制备了整体柱。整体柱表面的羟基经NaIO4氧化形成醛基后与胰蛋白酶的氨基进一步反应,实现胰蛋白酶的固定。在24s内,该酶反应器实现了肌红蛋白和细胞色素c的酶解,经MALDI-TOF MS鉴定,序列覆盖率分别达到65%和79%。

A monolithic enzymatic microreactor was prepared in a 500 μm fused-silica capillary by in situ polymerization of 2-hydroxyethyl methacrylate（HEMA） and ethylene dimethacrylate（EDMA） in the presence of a binary porogenic mixture of 1-decanol and cyclohexanol,followed by sodium periodate treatment and trypsin modification.The excellent performance of the monolithic microreactor was also demonstrated with the digestion of myoglobin and cytochrome c at the fast flow rate of 15 μL/min,which afforded a residence time of only 24 s.The digest was then characterized using MALDI-TOFMS,and the sequence coverage were 65% and 79%.