Use of chromatography to investigate biological recognition of a ligand byhuman serum albumin

Citation
Yc. Guillaume et al., Use of chromatography to investigate biological recognition of a ligand byhuman serum albumin, CHROMATOGR, 53(9-10), 2001, pp. 498-502
Citations number
42
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Chemistry & Analysis","Spectroscopy /Instrumentation/Analytical Sciences
Journal title
CHROMATOGRAPHIA
ISSN journal
0009-5893 → ACNP
Volume
53
Issue
9-10
Year of publication
2001
Pages
498 - 502
Database
ISI
SICI code
0009-5893(200105)53:9-10<498:UOCTIB>2.0.ZU;2-X
Abstract
Chromatographic temperature studies and differential scanning calorimetry h ave been used to elucidate the mechanism of retention of dansyl amino acids by human serum albumin (HSA) as stationary phase in reversed-phase liquid chromatography (RPLC) (Guillaume et al., Anal. Chem. 1997, 69, 4979). A nov el mathematical development is proposed for description of HSA-dansyl amino acid association when HSA is used as mobile-phase modifier and C-18 as sta tionary phase. The solute retention factor depends on the concentration of HSA in the bulk mobile phase and on the binding constant for the guest-HSA complex. The degree of complexation n(c) (the amount, %, of guest complexed ) was determined. Enthalpy-entropy compensation was also analyzed in relati on to this mathematical model to confirm the complexation behavior of the s olute with HSA. Variation of the binding constant with the concentration of sodium phosphate, i.e. with the electrostatic force controlling HA-solute association, is described. This theoretical model enabled estimation of the surface charge density(sigma /F) and the accessible solvent surface area o f the site II binding cavity; these were found to be approximately 8.7 x 10 (-7) molm(-2) and 2 nm(2).