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为了提高基因传递效率,本研究对用于基因传递的高分子/无机杂化纳米粒子进行了多肽功能化改性.利用生物素-亲和素(biotin-avidin)作用将同时具有肿瘤靶向性的精氨酸-甘氨酸-天冬氨酸(RGD)序列和细胞穿透功能的R8序列的生物素化多肽(biotinylated peptide,BP)修饰在具有良好生物相容性的生物素化肝素/鱼精蛋白/碳酸钙/DNA(HPB/PS/CaCO_3/DNA)杂化纳米粒子表面,通过水溶液中的自组装方法制备得到了BP/Avidin/HPB/PS/CaCO_3/DNA靶向基因传递纳米粒子.测量了其粒径和Zeta电位等,考察了该基因传递纳米粒子对HeLa细胞的转染效果.结果表明,BP/Avidin/HPB/PS/CaCO_3/DNA基因传递纳米粒子的水合粒径在175nm左右,Zeta电位为负值;与未经多肽功能化的传递系统相比,该纳米粒子能更好地被细胞摄入,达到更高的转染效率,说明多肽功能化能提高杂化纳米粒子的基因传递效果;与商品化的基因转染试剂Lipofectamine 2000相比,该纳米粒子介导荧光素酶(pGL3-Luc)质粒在细胞中的表达水平明显较高.
In order to improve the efficiency of gene transfer, we studied the functional modification of the polymer / inorganic hybrid nanoparticles used in gene delivery.Using the biotin-avidin method, The biotinylated peptide (BP) modification of the arginine-glycine-aspartic acid (RGD) sequence and the cell-penetrating functional R8 sequence was modified in a biocompatible biotinylated heparin / fish concentrate BP / Avidin / HPB / PS / CaCO_3 / DNA targeting gene delivery nanoparticles were prepared by self-assembly method in aqueous solution on the surface of HPA / HPB / PS / CaCO 3 / DNA hybrid nanoparticles. The particle size and Zeta potential of HepG2 cells transfected with HepG2 and HepG2 cells were investigated.The results showed that the hydrated particles of BP / Avidin / HPB / PS / CaCO 3 / DNA gene delivery nanoparticles were about 175 nm, Zeta potential is negative; compared with the delivery system without polypeptide functionalization, the nanoparticle can be better taken up by the cells to achieve higher transfection efficiency, indicating that polypeptide functionalization can increase the hybrid nanoparticle gene Delivery effect; and commercialization of gene transfection reagents Compared with Lipofectamine 2000, the expression level of the plasmid pGL3-Luc was significantly higher in the cells.