Enzymatic digestion of proteins is one of the key steps in proteomic analyses. There has been a steady progress in the applied digestion protocols in the past, starting from conventional time-consuming in-solution or in-gel digestion protocols to rapid and efficient methods utilizing different types of microscale enzyme reactors. Application of such microreactors has been proven beneficial due to lower sample consumption, higher sensitivity and straightforward coupling with LC-MS set-ups. Novel stationary phases, immobilization techniques and device formats are being constantly developed and tested to optimize digestion efficiency of proteolytic enzymes. This review focuses on the latest developments associated with the preparation and application of microscale enzyme reactors for proteomics applications since 2008 onwards. A special attention has been paid to the discussion of different stationary phases applied for immobilization purposes.
Keywords: ESI; Enzyme immobilization; Enzyme reactor; GNP; IMER; MOF; MSP; Microfluidics; Miniaturization; Monolith; NP; Proteomics; RPLC; electrospray ionization; gold nanoparticle; immobilized enzyme reactor; metal-organic framework; monolithic stationary phase; nanoparticle; reversed-phase liquid chromatography.
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