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On‐column derivatization of the antibiotics teicoplanin and ristocetin coupled to affinity capillary electrophoresis
Author(s) -
Silverio Catherine F.,
Azad Maryam,
Gomez Frank A.
Publication year - 2003
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/elps.200390101
Subject(s) - capillary electrophoresis , chromatography , teicoplanin , derivatization , chemistry , capillary electrochromatography , isoelectric focusing , antibiotics , high performance liquid chromatography , bacteria , biochemistry , biology , vancomycin , genetics , enzyme , staphylococcus aureus
Binding constants between the glycopeptides teicoplanin (Teic) and ristocetin (Rist) and their derivatives to D ‐Ala‐ D ‐Ala terminus peptides were determined by on‐column receptor synthesis coupled to partial‐filling affinity capillary electrophoresis (PFACE) or affinity capillary electrophoresis (ACE). In these techniques, the column is first partially filled with increasing concentrations of D ‐Ala‐ D ‐Ala terminus peptides. This is followed by plugs of buffer, antibiotic and two noninteracting standards, and acetic and/or succinic anhydride (and buffer in the case of ACE). The order of the reagent plugs containing the antibiotic and anhydride varies with the charge of the glycopeptide. Upon electrophoresis, the antibiotic reacts with the anhydride yielding a derivative of Teic or Rist. Continued electrophoresis results in the overlap of the derivatized antibiotic and the plug of D ‐Ala‐ D ‐Ala peptide. Analysis of the change in the relative migration time ratio (RMTR) of the new glycopeptide relative to the standards, as a function of the concentration of the D ‐Ala‐ D ‐Ala ligand yields a value for the binding constant K b . The techniques described here can be used to assess how the derivatization of drugs alters their affinities for target molecules.