title: A comparative study of capillary electrophoresis and isothermal titration calorimetry for the determination of binding constant of human serum albumin to monoclonal antibody.
abstract: This paper focuses on the investigation of the interactions between the anti-HSA-mAb and its protein antigen using CZE, ACE, and isothermal titration calorimetry. The CZE revealed the formation of the anti-HSA-mAb·HSA and anti-HSA-mAb·(HSA)2 complexes and the binding constants determined by plotting the amount of the bound anti-HSA-mAb as a function of the concentration of HSA. The ACE provided information on the binding strength from the change in effective electrophoretic mobility of the anti-HSA-mAb. These two separation techniques estimated the presence of two binding sites. The equilibrium dissociation constant values obtained by CZE and ACE were found to be 2.26 × 10(-6) M for anti-HSA-mAb·HSA, 1.22 × 10(-6) M for anti-HSA-mAb·(HSA)2 and 4.45 × 10(-8) M for anti-HSA-mAb·HSA, 1.08 × 10(-7) M for anti-HSA-mAb·(HSA)2 , respectively. The dissociation constant data obtained by ACE were in congruence with the values obtained by isothermal titration calorimetry (2.74 × 10(-8) M, 1.04 × 10(-7) M).
authors.affiliation: Department of Inorganic and Analytical Chemistry, University of Debrecen, Debrecen, Hungary.
authors.forename: Melinda
authors.initials: M
authors.lastname: Andrási
authors.affiliation: Department of Inorganic and Analytical Chemistry, University of Debrecen, Debrecen, Hungary.
authors.forename: Gábor
authors.initials: G
authors.lastname: Lehoczki
authors.affiliation: Department of Colloid and Environmental Chemistry, University of Debrecen, Debrecen, Hungary.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Nagy
authors.affiliation: Department of Inorganic and Analytical Chemistry, University of Debrecen, Debrecen, Hungary.
authors.forename: Gyöngyi
authors.initials: G
authors.lastname: Gyémánt
authors.affiliation: Department of Experimental Physics, University of Debrecen, Debrecen, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Pungor
authors.affiliation: Department of Inorganic and Analytical Chemistry, University of Debrecen, Debrecen, Hungary.
authors.forename: Attila
authors.initials: A
authors.lastname: Gáspár
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D001666:Binding Sites, Antibody; D002151:Calorimetry / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D012709:Serum Albumin / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D012709:Serum Albumin
keywords: ACE; Binding constant; CZE; HSA; mAb