Title
Rapid and enhanced proteolytic digestion using electric-field-oriented enzyme reactor Rapid and enhanced proteolytic digestion using electric-field-oriented enzyme reactor
Author
Faculty/Department
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences . Biomedical Sciences
Publication type
article
Publication
Subject
Chemistry
Biology
Source (journal)
Journal of proteomics
Volume/pages
74(2011) :7 , p. 1030-1035
ISSN
1874-3919
ISI
000292440400014
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Abstract
We have created a novel enzyme reactor using electric field-mediated orientation and immobilization of proteolytic enzymes (trypsin/chymotrypsin) on biocompatible PVDF membranes in a continuous flow-through chamber. Using less than 5 min, this reactor in various enzyme combinations can produce enhanced rapid digestion for standardized prototypic proteins, hydrophilic proteins and hydrophobic transmembrane proteins when compared to in-solution techniques. With improved digestive efficiency, our reactor improved the overall functional analysis of lipid raft proteomes by identifying more closely functionally linked proteins and elucidated a richer set of biological processes and pathways linked to the proteins,than traditional in-solution methods. Published by Elsevier B.V.
E-info
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