Characterization of Microorganisms and Biomarker Development from Global ESI-MS/MS Analyses of Cell Lysates

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• 作者：Fan Xiang ; Gordon A. Anderson ; Timothy D. Veenstra ; Mary S. Lipton ; and Richard D. Smith
• 刊名：Analytical Chemistry
• 出版年：2000
• 出版时间：June 1, 2000
• 年：2000
• 卷：72
• 期：11
• 页码：2475 - 2481
• 全文大小：180K
• 年卷期：v.72,no.11(June 1, 2000)
• ISSN：1520-6882

文摘

The capability for sensitive and accurate identification ofmicroorganisms has potential applications that includethe monitoring of industrial bioprocessing operations,food safety analyses, disease diagnosis, and detection ofpotential biological hazards. Efforts based upon matrix-assisted laser desorption/ionization mass spectrometryto detect and identify specific microorganisms have beenactively pursued for several years. We report a newmethod being developed to select useful biomarkers forthe identification of microorganisms based upon electrospray ionization (ESI)-ion trap mass spectrometry. Crudecell lysates are processed using a recently developed dual-microdialysis device and then directly infused into an iontrap MS. The low ESI flow rate and precursor ion accumulation capability of the ion trap MS enables high-sensitivity MS/MS analyses. Precursor ions are automatically selected and analyzed using tandem MS (MS/MS)to produce "global" MS/MS surveys and processed toyield two-dimensional MS/MS spectral displays. Suchglobal MS/MS surveys are demonstrated for Escherichiacoli lysates. The distinctive MS/MS spectral patterns canbe used to identify mass spectrometric-detected speciesuseful as biomarkers, which then provide a basis forconfident microorganism identification. The results presented demonstrate the application of this method for theidentification of microorganisms, as well as for detectionof bacteriophage MS2 in the presence of a large excess ofE. coli.