The Application Of Affinity Capillary Electrophoresis (ACE) In Drug-Protein Interaction Research

The affinity capillary electrophoresis (ACE) is a new area of capillary electrophoresis (CE). Beginning at the 90's, ACE has been widely used in the separating and analyses of biology samples such as protein, peptide, DNA or RNA fragments. ACE is often used for the study of interaction between molecules. The binding constant thus obtained can be used to elucidate the mode and strength of a given interaction, providing valuable information regarding molecules based on applications such as drug discovery and optimization.Because of the significant advantages and potential values in drug-protein interaction research and new compounds screening, ACE had been discussed deeply in this thesis. At first, BSA (Bovine Serum Albumin) and GT (Gatifloxacin) were choosen as model couples, two formats of ACE were investigated: one used GT as additive and the other one used BSA as additive. No literatures have studied the two formats using the BSA-GT model before. We study it from the initial experiment conditions. The value of binding constant between BSA and GT which calculated from the two formats matched well with each other. Besides, fluorescence method was used to validate the result. The result we measured from the experiments were quite according with those in the publishing literatures.Next, the use of ACE in the new drug screening was investigated. Two drugs: Glipizide and Amfebutamone were used in the experiment. Varying concentration of BSA was added in the running buffer, the two drugs were injected as simulant mixture sample. The result showed that both drugs interacted with BSA in one experiment as the peaks showed. At the same time Glipizide, Amfebutamone and their compounds seperated. This result hinted that ACE could be used in drug screening. More details need to be studied in the future experiments.