Structural changes of human serum albumin immobilized on chromatographic supports: a high-performance liquid chromatography and Fourier-transform infrared spectroscopy study

Chiral stationary phases obtained by immobilization of HSA on [C8] and [C18 ] reversed-phases and on poly(1-vinylimidazole)-coated silica were tested t o resolve DL-tryptophan, N-benzoyl-DL-phenylalanine, RS-oxazepam and RS-war farin racemic mixtures. Parameters of enantioselectivity measured in HPLC a re correlated to structural and solvation states for adsorbed HSA, evaluate d by FTIR spectroscopy. HSA immobilized on [PVI]-anion-exchangers is highly selective. HSA molecules are not self-associated, only unfolded for a smal l hydrophobic helix. The HSA-coated reversed-phases have a lower selectivit y. Unfolding is larger but the indole-benzodiazepine chiral site is preserv ed and remains accessible. (C) 2001 Elsevier Science B.V. All rights reserv ed.