将细胞固定在毛细管内壁上,利用亲和毛细管电泳研究受体配体相互作用并对组合化学合成化合物及天然产物进行筛选是近年来发展起来的新技术。在本文中,我们发展了一种将HEK293细胞固定在毛细管内壁上的新方法,对4个重要的固定化条件进行优化,其中包括细胞注入密度、PLL浓度、细胞培养时间和无菌处理方法。在优化后的实验条件下制备的固定化细胞贴壁毛细管柱,其均匀度、稳定性和耐用性良好,适用于毛细管电泳实验。该方法也可将超表达特定膜受体的HEK293细胞固定在毛细管内壁上,进而应用于靶向特定膜受体的亲和毛细管电泳药物筛选。
It is a new strategy to immobilize cells on the inner wall of a capillary column and use affinity capillary electrophoresis(ACE) to study receptor-ligand interactions or to screen natural products and compounds synthesized by combinatorial chemistry. In this paper, we developed a new method of immobilizing HEK293 cells on the inner wall of a capillary column. Four important experimental conditions were optimized, including cell injection density, PLL concentration, cell culturing time and sterile processing method. Immobilized cell-coated capillary columns prepared under the optimized experimental conditions exhibited good uniformity, stability and durability, which were suitable for capillary electrophoresis. The method could also be used to immobilize HEK293 cells over-expressing certain membrane receptors on the inner wall of a capillary. In this way, cell-coated capillary columns could be applied to ACE drug screening targeting certain membrane proteins.