Monolith‐based immobilized enzyme reactors: Recent developments and applications for proteome analysis

Abstract

In the current proteome study, protein digestion is indispensable before proteins could be identified by MS/MS, no matter based on top-down or bottom-up strategies. Compared to the traditional digestion performed in free solution, immobilized enzyme shows great advantages in digestion speed, stability, and longevity, especially with monolithic materials as the supports. Besides the improved digestion capacity, the immobilized enzyme reactors (IMERs) could be further coupled with the separation and detection systems, enabling high-throughput protein identification. In this paper, the latest advances in the monolith-based IMERs and their applications in proteomic study are briefly summarized. By reviewing these achievements, it could be seen that monilith-based IMERs have very bright future in proteome analysis.

Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis

“…As a large category of favorable supports for enzyme immobilization, monolithic supports have attracted significant attention due to facile preparation, fast mass transfer and low backpressure [18,19]. Generally, monolithic supports could be prepared by thermal polymerization [9][10][11], photopolymerization [20,21] and sol-gel technique [22][23][24], among which, photopolymerization has advantages of fast polymerization rate, controllable location and convenient operation both in UV-transparent capillaries and on microchips.…”

High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor

“…In response to these challenges, the immobilized enzyme reactor (IMER) [5][6][7][8] is a remarkable facility to perform digestion in a few minutes or even seconds. It could be also connected with related components directly for on-line digestion [9,10].…”

Integration of Biomolecules into analytical Systems by Means of Silica Sol-Gel Technology

“…Readers interested in the applications of monoliths to proteolytic enzyme reactors are advised to see the various comprehensive reviews. 12,66,67 To date, two techniques have been developed for immobilization of biomolecules on bimodal monolithic supports for LC: (1) the one-step immobilization of biomolecules on a bimodal monolith developed by Brennan et al and (2) the covalent bonding of biomolecules on a biomodal monolithic support. The latter technique is more popular than the former.…”