Research of interaction mechanism between human serum albumin and polyanions

HSA that has a affinity to a very wide range materials, including metals such as Cu and Zn, fatty acid, amino acid, metabolites such as bilirubin is used for drug delivery systems as a carrier to specific targets by a lot of researcher. In this thesis, interaction mechanism between HSA and PAA which is a polyanion is investigated and approach in bioengineering is improved for medicine, pharmacy, fundamental science and physico-chemical character of interaction is researched by using HPLC, GPC with quadruple dedector systems, 2+2+dynamic and electrophoretic light scattering and fluorescence techniques. Electrostatic interactions are the main forces in PPC formations; however, hydrophobic effects and hydrogen bondings are also possible. The interactions of polyelectrolytes (PE) with proteins in aqueous solutions is found to be pH, ionic strength and nHSA/nPAA ratio dependent. It was shown that the complex formation and the phase state of the systems depends on pH of the solution and ratio of components. For CPAA=1 mg/mL, Polyacrylic acid (PAA) and human serum albumine (HSA) form polymer-protein complexes which are water soluble or insoluble depending on pH values (pH=4.0-7.0) and protein/polymer ratios (nHSA/nPAA=0.1-10). The stability of PAA-HSA complexes at pH > pI is negligible weak (pH 7.0 and pH6,0), at the same time decreasing of pH (pH < pI) results with increasing of the stability of polycomplex particles and n/n dependencies of HSAPAAphysicochemical and conformational properties of of aqueous solutions of polycomplex particles at pH5,0 such as molecular properties and structural parameters (molecular weights (M, M), molecularweight distribution, radius of gyration (R), the Mark-Houwink equation constants (a), strenght of adsorption(k), saturated molar binding coefficient wnh(?m?)) is investigated. In addition, for CPAA=0,1 mg/mL pH effect to phase transition of policomplex particle at pH4,0 and pH4,5 is clarified using turbidimetric titration method. Key words: HPLC, GPC with quadruple dedector system, dynamic light scattering, electrophoretic light scattering, poly(acrylic acid), human serum albumin, protein-polyelectrolyte complex.