Capillary electrophoresis (CE) has many advantages, such as less reagent consumption, faster analysis and higher separation efficiency. It has been widely used to investigate the interaction of biomolecules. Liposome capillary electrophoresis (LCE) has been used to separate proteins, chiral molecules, and to investigate the interaction of bio-membrane and drugs. In this study, liposome was added into the buffers as pseudo-stationary phase (PSP), mimicking cell membrane. According to the establishment of biological circumstance in the capillary using capillary electrophoresis, the interactions of liposome-protein compounds with drugs could be studies, and the rapid screening model of drugs was established. After rotary evaporation, a dry lipid film could be achieved. Then sonication led to liposome suspension in which small spherical vesicles of lipid could be observed under microscope. Oxidation index was used to charactering inner-day stability of liposome, and the results showed that freeze-dried liposomes were more stable. The average size was 70 nm and the Zeta potential of liposome was -42.6mV, the liposome suspension was a relatively stable system. Then Carbonic Anhydrase (CA) was immobilized on liposomes by extrusion. A gradient concentrations of the immobilized enzyme were dispersed in Glycine-Tris (pH=8.5), as a pseudo-stationary phase. The binding constant of the 4-carboxy benzenesulfonamide, 4-chloro-3-sulfonyl-amino benzoic acid and 2, 4-dichloro-5-sulfonyl amino acid with liposome-CAB compound were 8.58�103(g/ml)-1, 9.06�103(g/ml)-1,1.20�104(g/ml)-1 respectively, and the drug screening model was established. Compared to the pure protein, the liposome-CA was much closer to the physiological environment. In our study, liposome-protein capillary electrophoresis can provide an important reference and basis for diuretic drugs.

Investigation of rapid drug screening model using liposome and Carbonic Anhydrase complex

245th National Meeting (2013)

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