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Istituto di scienza dell'alimentazione

Torna all'elenco Contributi in rivista anno 2005

Contributo in rivista

Tipo: Articolo in rivista

Titolo: Combination of solid-phase affinity capture on magnetic beads and mass spectrometry to study non-covalent interactions: example of minor groove binding drugs

Anno di pubblicazione: 2005

Formato: Elettronico Cartaceo

Autori: Malorni Antonio; Pocsfalvi Gabriella; Schlosser Gitta; Vékey Károly

Affiliazioni autori: Research Group of Peptide Chemistry - Hungarian Academy of Sciences; Proteomic and Biomolecular Mass Spectrometry Center - Institute of Food Science and Technology; Proteomic and Biomolecular Mass Spectrometry Center (CeSMa-ProBio) - Institute of Food Science and Technology; Institute of Structural Chemistry - Hungarian Academy of Sciences

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Lingua: inglese

Abstract: A simple and novel approach was developed to detect non-covalent interactions. It is based on combination of solid-phase affinity capture with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). One of the interacting molecules is bound to magnetic beads and is incubated with the target molecules in solution. The complex bound on the solid support is removed from the solution and transferred for MALDI analysis. Mass spectrometry is used only to detect the target compound, which is far more straightforward than detecting the intact non-covalent complex. To demonstrate the applicability of the method, an AT-rich oligonucleotide (5'-CCCCCAATTCCCCC-3') and its complementary biotinylated sequence (5'-biotin-GGGGGAATTGGGGG-3') were hybridized and immobilized to paramagnetic particles by streptavidin-biotin interaction. The immobilized duplex oligonucleotide was reacted with minor groove binding drugs, Netropsin, Distamycin A, Hoechst 33258 and 4',6-diamidino-2-phenylindole. The resulting DNA-drug complex bound to the particles was separated and analyzed by linear MALDI-TOFMS after washing. Drugs were selectively detected in the spectra. Relative binding strengths were also estimated using competitive complexation. Copyright (c) 2005 John Wiley & Sons, Ltd.

Lingua abstract: inglese

Pagine da: 3307

Pagine a: 3314


RCM. Rapid communications in mass spectrometry Heyden,
Paese di pubblicazione: Regno Unito
Lingua: inglese
ISSN: 0951-4198

Numero volume: 19

Numero fascicolo: 22

DOI: 10.1002/rcm.2193

Referee: Sě: Internazionale

Indicizzato da: ISI Web of Science (WOS) [000233483500019]

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