Subject Area物理化学
Novel capillary and microchip based enzymatic reactors with hydrophilic monolithic polymer as matrix
Liang Y(梁玉); Ma JF(马俊锋); Liang Z(梁振); Zhang LH(张丽华); Zhang YK(张玉奎)
Conference NameThe 24th MSB 2009
Conference Date2009-10-19
Conference Place中国
Funding Organization大连化物所
AbstractDigestion is an indispensable step for protein analysis with bottom-up strategy. However, the traditional in solution protein digestion has unavoidable shortcomings, such as long digestion time, unavoidable enzyme autodigestion, and manual operation. To resolve these problems, microscale immobilized enzyme reactors, prepared either in capillaries or microfluidic channels, are highly desirable for minute protein analysis with improved digestion capacity and throughput. Particles and monolithic supports have been widely used for enzyme immobilization [1, 2]. Compared to particulate supports, monoliths have obtained much attention due to ease-in-preparation, large surface area, fast mass transfer and low back pressure. However, nonspecific adsorption of proteins onto such supports is still a serious problem [3]. In this study, a novel kind of enzymatic reactor with hydrophilic monolith as support was prepared in UV-transparent capillaries and microchip channels by the photo initiated polymerization of N-Acryloxysuccinimide and poly (ethylene glycol) diacrylate, followed by trypsin immobilization. Since succinimide groups are readily to react with amine groups of trypsin, the whole preparation procedure is quite facile. More importantly, the monolithic support shows good hydrophilicity, which could avoid the undesired nonspecific adsorption of proteins and /or peptides. The performance of such enzymatic microreactor was demonstrated by the digestion of myoglobin. With digests analyzed by μRPLC-MS/MS, the yielding sequence coverage for on-column digestion within 3 min was higher than that obtained by in-solution digestion within 12h. Moreover, the microreactor was successfully applied to the digestion of proteins extracted from E. coli.
Document Type会议论文
Corresponding AuthorZhang LH(张丽华)
Recommended Citation
GB/T 7714
Liang Y,Ma JF,Liang Z,et al. Novel capillary and microchip based enzymatic reactors with hydrophilic monolithic polymer as matrix[C],2009:246/1.
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