Electrophoretic Screening Of Enzymatic Activity And Inhibitors And Study Of Wide-bore Electrophoretic Technique

As being a developing project, drug screening has been attracted increased attention since coming into being due to its both clinic value and social significance. In recent years, more interests are focusing on the improvement of efficiency and accuracy of screening methods to deal with the plenty of potential drug candidates. Capillary electrophoresis (CE) is widely perceived as a fast, economical, and efficient separation technique and especially, when used in combination with laser-induced fluorescence (LIF) detection, has high sensitivity. This separation-based technique can compensate the shortage of some traditional drug screening methods. On the other hand, the study of novel candidates which specially target the special enzyme is an important aspect in the pharmaceutical industry. Herein, we present a robust CE/LIF method, using tyrosine kinase and signal peptides as probe enzymes, and systematically proved the feasibility of CE method performing the characterization and inhibitor screen.Meanwhile, some recent improvements have been reported for enhancing the loading capacity of CE-based methods, however, loading capacity is still a critical issue if CE approaches are to be employed in multi-dimensional separation approaches. Based on previous reports, a novel wide-bore (WE) electrophoresis method employing nanoparticles as a pseudostationary phase was evaluated for improving separation efficiency using a model mixture of six aromatic acids. The nanoparticles used in this work were prepared in-house by an electrospray method. The results indicted that the sample loading capacity and resolution were both significantly increased by utilizing the novel WE approach in conjunction with aminopropyl modified nanoparticles as an additive in running buffer.In summary, traditional CE methods is an operational method which may contribute to the improvement of drug screen and high throughput screen methods. And the exploration and optimization of novel wide-bore electrophoresis enhanced the practicability and widen the field of electrophoresis techinique.