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On‐probe sample pretreatment for the detection of proteins above 15 KDa from whole cell bacteria by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Madonna Angelo J.,
Basile Franco,
Ferrer Imma,
Meetani Mohammed A.,
Rees Jon C.,
Voorhees Kent J.
Publication year - 2000
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/1097-0231(20001215)14:23<2220::aid-rcm155>3.0.co;2-4
Subject(s) - chemistry , formic acid , mass spectrometry , chromatography , matrix assisted laser desorption/ionization , bacteria , desorption , matrix (chemical analysis) , sample preparation , surface enhanced laser desorption/ionization , time of flight mass spectrometry , ferulic acid , analytical chemistry (journal) , ionization , protein mass spectrometry , tandem mass spectrometry , organic chemistry , ion , adsorption , biology , genetics
A rapid methodology is described for the enhancement of the signal‐to‐base‐line (S/B) ratio of high molecular weight protein signals from whole cell bacteria analyzed by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS). The procedure involves depositing growing bacteria colonies from culture dishes directly onto the MALDI probe followed by treatment of the sample spot with a 2 µL aliquot of 40% ethanol prior to the addition of a ferulic acid matrix solution (12.5 mg dissolved in 17% formic acid/33% acetonitrile/50% H 2 O). Protein signals of more than 20 kDa were routinely produced from both Gram positive and Gram negative bacteria prepared in this manner. Moreover, a substantial number of intense protein signals were also produced in the more ‘ conventional ’ fingerprint region extending from 4 to 20 kDa. This approach is rapid, easy to implement into existing methodologies, and does not require any special hardware. Copyright © 2000 John Wiley & Sons, Ltd.