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/lp/wiley/on-line-protein-digestion-and-peptide-mapping-by-capillary-k5EmZLChxJ
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- Publisher
- Wiley
- Copyright
- "Copyright © 2004 Wiley Subscription Services, Inc., A Wiley Company"
- ISSN
- 0173-0835
- eISSN
- 1522-2683
- DOI
- 10.1002/elps.200305841
- pmid
- 15174054
- Publisher site
- See Article on Publisher Site
Abstract
A nanoliter enzyme microreactor was developed for on‐line capillary electrophoresis (CE) peptide mapping of proteins, allowing picomole quantities of proteins to be digested. The enzyme microreactor was formed by immobilizing trypsin onto a monolithic capillary column, which was prepared by in situ polymerization of glycidyl methacrylate and ethylene dimethacrylate in a capillary. Highly efficient digestion of three protein standards was demonstrated. The detection of peptide fragments in CE was enhanced by post‐column derivatization and laser‐induced fluorescence detection. The microreactor has a volume of about 30 nL and is coupled with a separation capillary via a fluid joint for on‐line digestion. The overall analysis, including digestion and separation, lasted only about 16 min. Column efficiencies > 300 000 plates/m were obtained for most peaks in the electropherogram of an on‐line peptide mapping experiment of denatured α‐lactalbumin under optimal conditions.
Journal
Electrophoresis – Wiley
Published: May 1, 2004
Keywords: ; ; ;