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Rapid protein identification using a microscale electrospray LC/MS system on an ion trap mass spectrometer

  • Michael T. Davis
  • Terry D. LeeEmail author
Articles

Abstract

A methodology has been developed for the rapid identification of gel separated proteins. Following in gel protein digestion with trypsin, the resulting peptide mixture is analyzed by on-line liquid chromatography, electrospray mass spectrometry (LC/MS). The mass spectral data containing either accurate mass values or sequence specific fragment ion information is then matched to a database of known protein sequences. Key features of the LC/MS system are the use of a novel integrated, microscale LC column-electrospray interface and variable flow solvent delivery to optimize the efficiency of sample loading and gradient elution. With these enhancements, only 10 min is required to analyze each sample. The method is routine for sample amounts ranging from 50 to 500 fmol. The analysis parameters for the ion trap mass spectrometer have to be carefully adjusted in order to keep pace with the rapidly eluting LC peaks. Although designed for rapid LC separations, the integrated column-electrospray interface is also able to provide extended analyses of selected components using a technique known as “peak parking. ”

Keywords

Collision Induce Dissociation Fuse Silica Capillary Automatic Gain Control Mass Spectral Analysis Base Peak Chromatogram 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© American Society for Mass Spectrometry 1998

Authors and Affiliations

  1. 1.Division of ImmunologyBeckman Research Institute of the City of HopeDuarte

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