ESTIMATION OF THE NUMBER OF ENANTIOSELECTIVE SITES OF BOVINE SERUM-ALBUMIN USING FRONTAL CHROMATOGRAPHY

On a column with bovine serum albumin (BSA) immobilized covalently to silica, the adsorption isotherms of the enantiomers of mandelic acid, tryptophan, 2-phenylbutyric acid, and N-benzoylalanine are measured us ing a buffered mobile phase. Knowing the amount of BSA immobilized on the column (36 mg), the ratio of the number of enantiomer molecules ne eded to saturate the enantioselective retention mechanism to the numbe r of BSA molecules is determined. The mean of the set of eight enantio mers is 0.28. These data confirm that at most one enantioselective sit e exists for each BSA molecule for the kind of enantiomers studied. (C ) 1993 Wiley-Liss, Inc.