title: The automatic use of capillary isoelectric focusing with whole column imaging detection for carbamazepine binding to human serum albumin.
abstract: The binding of the anticonvulsant drug carbamazepine (CBZ) to human serum albumin, both without (dHSA) and in the presence of fatty acids (HSA) was studied in real time by capillary isoelectric focusing with whole column imaging detection (cIEF-WCID). Reaction mixtures at different CBZ:HSA and CBZ:dHSA molar ratios (0:1/25:1) were prepared in phosphate buffer saline (PBS) solution at a physiological pH (7.4), and incubated for 0-72h at 37°C in a water bath. Application of the cIEF-WCID method allowed for observations on the impact of increasing CBZ:serum albumin molar ratios on isoelectric point (pI) shifts, as well as changes in peak area and absorbance, which serve as evidence of structural alterations occurring in the protein in the presence of CBZ. The obtained cIEF-WCID results indicated that the dynamic process of complex formation is not dependent on incubation time. The presented work allowed for recognition of different types of interactions, as well as for the calculation of association constants that demonstrate the stability of the complex. This study was also designed to examine the possible impact of fatty acids (FAs) on protein stability and drug delivery in blood.
authors.affiliation: Department of Chemistry, University of Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Maciążek-Jurczyk
authors.affiliation: Department of Chemistry, University of Waterloo, Ontario N2L 3G1, Canada. Electronic address: janusz@uwaterloo.ca.
authors.forename: Janusz
authors.initials: J
authors.lastname: Pawliszyn
mesh_terms: D000927:Anticonvulsants / Q000097:blood*; D002021:Buffers; D002220:Carbamazepine / Q000097:blood*; D005227:Fatty Acids / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007525:Isoelectric Focusing / Q000379:methods*; D011485:Protein Binding; D055550:Protein Stability; D012709:Serum Albumin / Q000378:metabolism*
chemical_list: D000927:Anticonvulsants; D002021:Buffers; D005227:Fatty Acids; D012709:Serum Albumin; D002220:Carbamazepine
keywords: Carbamazepine; Fatty acids; Human serum albumin; Isoelectric point; cIEF-WCID