Premium
Rapid determination of protein binding constant by a pressure‐mediated affinity capillary electrophoresis method
Author(s) -
Zhang Bo,
Li You X.,
Gao He N.,
Bian Juan,
Bao James J.
Publication year - 2011
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.201100211
Subject(s) - capillary electrophoresis , affinity electrophoresis , chemistry , electrophoresis , fluorescence spectroscopy , binding constant , chromatography , fluorescence , bovine serum albumin , gel electrophoresis of proteins , analytical chemistry (journal) , binding site , polyacrylamide gel electrophoresis , affinity chromatography , enzyme , biochemistry , physics , quantum mechanics
A new pressure‐mediated affinity capillary electrophoresis method for the rapid and accurate determination of drug–protein binding constants is described. A special combination of pressure and electrophoresis is used to shorten the electrophoresis and the overall analysis time to only a few minutes. At the same time, the suitability of this method is checked against a traditional fluorescence spectroscopy method. The binding constants of bovine serum albumin and a total of eight drugs with different p K a have been evaluated and compared with those determined by the fluorescence spectroscopy method and other methods in literature. The results indicate that the P‐ACE method is well suited for the determination of binding constants with weak interaction ( K b <10 5 M −1 ).