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A novel organic-inorganic hybrid monolith for trypsin immobilization

机译：一种新型的固定化胰蛋白酶的有机-无机杂化整体

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摘要

In proteomics, attention has focused on various immobilized enzyme reactors (IMERs) for the realization of high throughput digestion. In this report, a novel organic-inorganic hybrid monolith based IMER was prepared in a 100 μm i.d. capillary with 3-glycidoxypropyltrimethoxysilane (GLYMO) as the monomer and tetraethoxysilane (TEOS) as the crosslinker. Trypsin immobilization was achieved via the reaction between vicinal diol groups, which were obtained from hydrolysis of epoxy groups, and the amino groups of trypsin. Bovine serum albumin was digested thoroughly by this IMER in 47 s. After micro-reverse phase liquid chromatography-tandem mass spectrometry (μRPLC- MS/MS ) analysis and database searching, beyond 35% sequence coverage was obtained, and the result was comparable to that of 12 h in solution digestion. The present IMER has potential for high throughput digestion.
机译：在蛋白质组学中，注意力集中在实现高通量消化的各种固定化酶反应器（IMER）上。在本报告中，以100μmi.d.制备了一种新型的基于有机-无机杂化单体的IMER。以3-环氧丙氧基丙基三甲氧基硅烷（GLYMO）为单体，四乙氧基硅烷（TEOS）作为交联剂的毛细管。通过环氧基的水解获得的邻二醇基团与胰蛋白酶的氨基之间的反应实现了胰蛋白酶的固定化。该IMER在47 s内彻底消化了牛血清白蛋白。经微反相液相色谱-串联质谱（μRPLC-MS/ MS）分析和数据库搜索，获得了超过35％的序列覆盖率，其结果与溶液消化的12 h相当。本IMER具有高通量消化的潜力。

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