A replaceable microreactor for on-line protein digestion in a two-dimensional capillary electrophoresis system with tandem mass spectrometry detection |
| |
Authors: | Li Yihan Wojcik Roza Dovichi Norman J |
| |
Institution: | Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA. |
| |
Abstract: | 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 β-casein as model proteins. |
| |
Keywords: | Two-dimensional capillary electrophoresis ESI-MS Magnetic beads |
本文献已被 ScienceDirect PubMed 等数据库收录! |