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Considerations concerning interaction characterization of oligopeptide mixtures with vancomycin using affinity capillary electrophoresis‐electrospray mass spectrometry
Author(s) -
Lynen Frédéric,
Zhao Yining,
Becu Christian,
Borremans Frans,
Sandra Pat
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990801)20:12<2462::aid-elps2462>3.0.co;2-5
Subject(s) - capillary electrophoresis , chemistry , mass spectrometry , electrospray ionization , dissociation constant , chromatography , capillary electrophoresis–mass spectrometry , dissociation (chemistry) , electrospray , electrophoresis , electrolyte , analytical chemistry (journal) , combinatorial chemistry , receptor , biochemistry , electrode
In the past few years affinity capillary electrophoresis (ACE) has proven to be a powerful tool to study molecular interactions. In ACE the change in electrophoretic mobility between a free and a complexed ligand with a receptor dissolved in the background electrolyte is observed. It provides an accurate way to calculate binding or dissociation constants and, when coupled to mass spectrometry, it forms a promising method to analyze solution‐based combinatorial libraries. We report a model study on the macrocyclic antibiotic vancomycin using a 36‐component library of tetrapeptides of the type 9‐fluorenylmethoxycarbonyl (Fmoc)‐ L ‐Asp‐ L ‐Asp‐ D ‐Xaa‐ D ‐Xaa. The mass spectrometry conditions were optimized by fine‐tuning the background electrolyte and sheath flow composition to achieve optimal sensitivity in the negative ionization mode. Different types of capillaries were also evaluated on their potential to screen combinatorial libraries. The library components that show the strongest interaction were identified. The dissociation constants of a mixture of six compounds with a broad affinity range were simultaneously established by Scatchard analysis on ACE‐MS.