期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1218, 期 15, 页码 2007-2011出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2010.10.013
关键词
Two-dimensional capillary electrophoresis; ESI-MS; Magnetic beads
资金
- National Institutes of Health [R33CA122900]
We describe a two-dimensional capillary electrophoresis system that incorporates a replaceable enzymatic microreactor for on-line protein digestion. In this system, trypsin is immobilized on magnetic beads. At the start of each experiment, old beads are flushed to waste and replaced with a fresh plug of beads, which is captured by a pair of magnets at the distal tip of the first capillary. For analysis, proteins are separated in the first capillary. A fraction is then parked in the reactor to create peptides. Digested peptides are periodically transferred to the second capillary for separation; a fresh protein fraction is simultaneously moved to the reactor for digestion. An electrospray interface is used to introduce peptides into a mass spectrometer for analysis. This procedure is repeated for several dozen fractions under computer control. The system was demonstrated by the separation and digestion of insulin chain b oxidized and beta-casein as model proteins. (C) 2010 Elsevier B.V. All rights reserved.
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