A comparative study of capillary electrophoresis and isothermal titration calorimetry for the determination of binding constant of human serum albumin to monoclonal antibody

作者：Andrasi Melinda^*; Lehoczki Gabor; Nagy Zoltan; Gyemant Gyoengyi; Pungor Andras; Gaspar Attila

来源：Electrophoresis, 2015, 36(11-12): 1274-1281.

DOI：10.1002/elps.201400513

• ACE
• Binding constant
• CZE
• HSA
• mAb

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 center dot HSA and anti-HSA-mAb center dot(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 x 10(-6) M for anti-HSA-mAb center dot HSA, 1.22 x 10(-6) M for anti-HSA-mAb center dot(HSA)(2) and 4.45 x 10(-8) M for anti-HSA-mAb center dot HSA, 1.08 x 10(-7) M for anti-HSA-mAb center dot(HSA)(2), respectively. The dissociation constant data obtained by ACE were in congruence with the values obtained by isothermal titration calorimetry (2.74 x 10(-8) M, 1.04 x 10(-7) M).

• 出版日期2015-6