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

Measurement of drug–protein binding by immobilized human serum albumin-HPLC and comparison with ultrafiltration

“…The earliest work in the area of human serum immobilized chromatography was carried out by Wainer and his group [34][35][36][37]. Henceforth, different strategies for the development of columns immobilized with HSA have been reported [38][39][40][41][42][43][44]. In one method, the epoxy groups of this copolymer (glycidyl methacrylate and ethylene dimethacrylate) were used directly for the immobilization of HSA through its amine residues (i.e., the epoxy method); in other approaches, these epoxy groups were converted to diols for later use in the carbonyldiimidazole, disuccinimidyl carbonate, and Schiff base methods [38].…”

“…Human serum albumin (HSA) has been reported to be successfully bonded to silica with N-hydroxysuccinimide [39] and s-triazine as activator [44]. Stationary phases obtained by immobilization of HSA on [C8] and [C18] reversed-phases and on poly(1-vinylimidazole)-coated silica have also been reported [41,43].…”

Separation of drug enantiomers by liquid chromatography and capillary electrophoresis, using immobilized proteins as chiral selectors

“…The immobilization procedure resulted in a stable protein layer since no leakage of BSA was detected for at least 1 week. Chiral separations were observed under suitable elution conditions, even if adsorption techniques based on hydrophobic interactions between the protein and the support may entail significant conformational changes for the biomolecule, as it was shown recently by FTIR for HSA molecules adsorbed on reversed-phase silica particles [69].…”

“…In a recent study, HSA was adsorbed to quaternized poly(vinylimidazole)-coated silica particles, resulting in performant chiral stationary phases [67]. It was shown by Fourier-transform infrared spectroscopy (FTIR) that the adsorbed HSA molecules were only unfolded in a small hydrophobic helix corresponding to 3% of the backbone [69]. Thus, the ionic adsorption of proteins to ion-exchange matrices can be considered as a gentle method resulting in minor structural changes.…”

Effect of morphine, oxycodone and thebaine on resonance light scattering properties of human serum albumin: Investigation possibility of morphine determination in the presence of the two other drugs

“…Therefore, it has become an important research field in chemistry, life science, and clinical medicine. Some techniques commonly used to detect interaction between drug and serum albumin include the fluorescence spectroscopy [4], UV-spectrophotometry [5], circular dichroism spectroscopy [6], Raman spectroscopy [7], nuclear magnetic resonance [8], electrochemistry [9,10], equilibrium dialysis [11] and attenuated total reflectance-Fourier transform infrared [12].…”