Protein fingerprinting of staphylococcus aureus by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection

  1. Pelaez-Lorenzo, C. 12
  2. Veledo, M.T. 24
  3. Gonzalez, R. 3
  4. De Frutos, M. 2
  5. Diez-Masa, J.C. 2
  1. 1 Universidad Europea de Madrid
    info

    Universidad Europea de Madrid

    Madrid, España

    ROR https://ror.org/04dp46240

  2. 2 Instituto de Química Orgánica General
    info

    Instituto de Química Orgánica General

    Madrid, España

    ROR https://ror.org/05e0q7s59

  3. 3 Instituto de Ciencias de la Vid y del Vino
    info

    Instituto de Ciencias de la Vid y del Vino

    Logroño, España

    ROR https://ror.org/01rm2sw78

  4. 4 PharmaMar, Madrid, Spain
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Zeitschrift:
Methods in molecular biology

ISSN: 1064-3745

Datum der Publikation: 2013

Ausgabe: 984

Seiten: 237-251

Art: Artikel

DOI: 10.1007/978-1-62703-296-4-17 PMID: 23386348 SCOPUS: 2-s2.0-84880145233 GOOGLE SCHOLAR

Andere Publikationen in: Methods in molecular biology

Zusammenfassung

This chapter describes a complete procedure for obtaining protein fingerprints of microorganisms using capillary electrophoresis (CE) with laser-induced fluorescence detection (LIF). Staphylococcus aureus, a human pathogen responsible of frequent and resistant infections, is used as model microorganism to show the feasibility of this procedure. Bacteria are grown in different culture media or submitted to temperature or nitrosative stress conditions. After the growth of the bacteria, the protein extracts are obtained by cell lysis using sonication. The water-soluble fraction of these lysates is derivatized on-capillary with the fluorogenic reagent 3-(2-furoyl)quinoline-2- carboxaldehyde. The fluorescent products are analyzed by CE and detected by LIF. Practical advices for the interpretation of the electropherograms are given. To do so, the variations of the protein fingerprints of the bacteria with the culture conditions, such as growth medium, or the stressing conditions, such as heat shock or nitrosative stress, are used as example. © Springer Science+Business Media, LLC 2013.