Use of Frontal Chromatography to Measure the Binding Interaction of Berberine Chloride with Bovine Serum Albumin
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There is much interest in the interactions between the active constituents of traditional Chinese medicine and biomolecules. By use of frontal analysis on an affinity column we have examined the binding interaction of berberine chloride (BC), a major active constituent of coptis, with bovine serum albumin (BSA) in 40 mM phosphate buffer, pH 7.0. Adsorption of BC on immobilized BSA was in accordance with the Langmuir isotherm, suggesting BC is binding to a single type of site on the immobilized BSA. The binding constant was 4.79 × 104 L mol−1 at 30 °C, less than the value of 6.61 × 104 L mol−1 obtained by fluorescence spectroscopy under the same buffer and temperature conditions. The effects of temperature on the retention, binding constant, and active binding sites, and on the percentage protein binding of BC, were also investigated. Thermodynamic measurements indicated that the increase in entropy was an important process promoting the interaction between BC and BSA.
KeywordsAffinity chromatography Binding interaction Bovine serum albumin Berberine chloride
This work was supported by grants from the National Natural Science Foundation of China (No. 20575052), the Natural Science Foundation of Shaanxi Province (No. 2006B03), and the Science Foundation of Northwest University (No. 04NW42).
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