Chromatographic Chiral Separations on Immobilised Proteins: Does High-Performance Liquid Chromatography Mirror Nature?
A commercially available high-performance liquid chromatography (HPLC) chiral stationary phase (CSP) based upon immobilised bovine serum abumin, the BSA-CSP, has been used to investigate the protein binding of enantiomeric solutes. The changes in the chromatographic retention which resulted from the addition of mobile phase modifiers were compared with parallel in vitro protein binding studies. The results of these studies indicate that the commercially available BSA-CSP can be used to investigate the binding interactions between a series of solutes and the protein. The use of mobile phase modifiers has demonstrated a direct relationship between in vitro protein binding and chromatographic retention. In addition, the use of enantiomeric solutes has shown that the BSACSP can be used to probe the number and relative affinities of various binding sites on the protein.
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