Determination of Bacterial Protein Profiles by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry with High-performance Liquid Chromatography

Determination of Bacterial Protein Profiles by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry with High-performance Liquid Chromatography

Determination of Bacterial Protein Profiles by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry with High-performance Liquid Chromatography

Abstract

A rapid method for profiling bacterial and cellular proteins has been developed using a combination of capillary high-performance liquid chromatography separation followed by (MALDI-MS) matrix-assisted laser desorption/ionization mass spectrometric analysis. In this method, bacteria are sonicated, the cell walls broken, and the water-soluble proteins precipitated for analysis. The proteins are separated by capillary liquidchromatography and detected on-line by a UV absorption detector. The eluents are then collected for off-line analysis by MALDI-MS. Using this method, it is demonstrated that bacteria can be discriminated based upon their protein profiles to the species level with only pmol level detection ofproteins. It has also proved to be a fast and accurate means for monitoring the expression of Hsp27 in an insect cell system.

read online

Post Comment