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Istituto di scienza dell'alimentazione

Torna all'elenco Contributi in rivista anno 2007

Contributo in rivista

Tipo: Articolo in rivista

Titolo: Coupling Immunomagnetic Separation on Magnetic Beads with MALDI-TOF Mass Spectrometry for the Detection of Staphylococcal Enterotoxin B

Anno di pubblicazione: 2007

Formato: Elettronico Cartaceo

Autori: Schlosser Gitta; Kacer Petr; Kuzma Marek; Szilagyi Zoltan; Sorrentino Alida; Manzo Carla; Pizzano Rosa; Malorni Livia; Pocsfalvi Gabriella Katalin

Affiliazioni autori: Malorni Livia, Manzo Carla, Pizzano Rosa, Pocsfalvi Gabriella Katalin, Sorrentino Alida: Istituto di scienza dell'alimentazione; Kacer Petr: Department of Organic Technology - Institute of Chemical Technology, Prague, Czech Republic; Schlosser Gitta: Research Group of Peptide Chemistry, Department of Organic Chemistry - Hungarian Academy of Sciences, Eötvös L. University, Budapest, Hungary; Kuzma Marek: Institute of Microbiology - Academy of Sciences of the Czech Republic, Prague, Czech Republic; Szilagyi Zoltan: Research Institute for Viticulture and Enology, Kolyukteto, H-3300, Eger, Hungary

Autori CNR:

  • LIVIA MALORNI
  • ROSA PIZZANO
  • GABRIELLA KATALIN POCSFALVI
  • ALIDA SORRENTINO

Lingua: inglese

Abstract: The growing importance of mass spectrometry for the identification and characterization of bacterial protein toxins is a consequence of the improved sensitivity and specificity of mass spectrometry-based techniques, especially when these techniques are combined with affinity methods. Here we describe a novel method based on the use of immunoaffinity capture and matrix-assisted laser desorption ionization-time of flight mass spectrometry for selective purification and detection of staphylococcal enterotoxin B (SEB). SEB is a potent bacterial protein toxin responsible for food poisoning, as well as a potential biological warfare agent. Unambiguous detection of SEB at low-nanogram levels in complex matrices is thus an important objective. In this work, an affinity molecular probe was prepared by immobilizing anti-SEB antibody on the surface of para-toluene-sulfonyl-functionalized monodisperse magnetic particles and used to selectively isolate SEB. Immobilization and affinity capture procedures were optimized to maximize the density of anti-SEB immunoglobulin G and the amount of captured SEB, respectively, on the surface of magnetic beads. SEB could be detected directly "on beads" by placing the molecular probe on the matrix-assisted laser desorption ionization target plate or, alternatively, "off beads" after its acidic elution. Application of this method to complex biological matrices was demonstrated by selective detection of SEB present in different matrices, such as cultivation media of Staphylococcus aureus strains and raw milk samples.

Lingua abstract: inglese

Pagine da: 6945

Pagine a: 6952

Pagine totali: 8

Rivista:

Applied and environmental microbiology American Society for Microbiology.
Paese di pubblicazione: Stati Uniti d'America
Lingua: inglese
ISSN: 0099-2240

Numero volume: 73

Numero fascicolo: 21

DOI: 10.1128/AEM.01136-07

Indicizzato da:

  • ISI Web of Science (WOS) [000250700600029]
  • Scopus [2-s2.0-35948957858]
  • PubMed [17827336]

Parole chiave:

  • affinity capture
  • immunomagnetic separation
  • staphylococcus aureus
  • MALDI-TOF
  • Staphylococcal enterotoxin B

Strutture CNR:

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