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

Gitta Schlosser, Károly Vékey, Antonio Malorni, Gabriella Pocsfalvi

Research output: Contribution to journalArticle

16 Citations (Scopus)

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-phenyl-indole. 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.

Original languageEnglish
Pages (from-to)3307-3314
Number of pages8
JournalRapid Communications in Mass Spectrometry
Volume19
Issue number22
DOIs
Publication statusPublished - 2005

Fingerprint

Mass spectrometry
Biotin
Oligonucleotides
Ionization
Desorption
Netropsin
Pharmaceutical Preparations
Bisbenzimidazole
Molecules
Streptavidin
Lasers
Complexation
Washing
DNA
stallimycin
indole

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy

Cite this

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title = "Combination of solid-phase affinity capture on magnetic beads and mass spectrometry to study non-covalent interactions: Example of minor groove binding drugs",
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-phenyl-indole. 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.",
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AU - Malorni, Antonio

AU - Pocsfalvi, Gabriella

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