载基因阳离子脂质体的制备与包封率测定  

Preparation of gene-loaded cationic liposomes and the determination of entrapment efficiency

作　　者：杨硕晔[1] 郑义[1] 陈佳音[1] 赵娣[1] 陈西敬[1] 

机构地区：[1]中国药科大学药物代谢动力学研究中心,南京210009

出　　处：《中国新药杂志》2011年第20期2030-2034,共5页Chinese Journal of New Drugs

摘　　要：目的:制备包封基因质粒的阳离子脂质体并考察其性质、测定包封率。方法:以DC-Chol和DOPE为材料,薄膜分散法制备阳离子脂质体,与可表达增强型绿色荧光蛋白的基因质粒结合并考察其转染性能,激光粒度分析仪测定阳离子脂质体和脂质复合物的粒径及Zeta电位;使用葡聚糖凝胶过滤法测定包封率,并对该法进行详细考察。结果:所制备的阳离子脂质体和脂质复合物的平均粒径分别为161.6和216.3 nm,Zeta电位分别为+22.2和+3.2 mV;基因质粒在0.1925～3.85μg.mL-1浓度范围内线性良好,精密度高,与葡聚糖无吸附作用,柱回收率高,测得脂质复合物的包封率为89.94%。结论:采用该处方和工艺可成功制备质量良好、能有效转染细胞的阳离子脂质体载体,葡聚糖凝胶过滤法可准确测定其包封率,该法快速、简便、有效。Objective： To prepare gene-loaded cationic liposomes,to evaluate the properties,and to determine the entrapment efficiency.Methods： Cationic liposomes were mixed with the plasmid DNA that can express green fluorescent protein（GFP）,and prepared by film dispersion method with DC-Chol and DOPE.The transfection experiment was conducted subsequently.Laser particle analyzer was applied to determine the size and Zeta potentials of cationic liposomes and lipoplexes.Free DNA and liposomes were separated by sephadex column and the entrapment efficiency was determined using an ultraviolet spectrophotometer.Results： The mean diameters were 161.6 and 216.3nm,and Zeta potentials were ＋22.2 and ＋3.2mV of cationic liposomes and lipoplexes,respectively.Plasmid DNA did not be absorbed by Sephadex G-50.A good linear relationship was found in the rages of 0.1925~3.85μg·mL-1（r=0.9988,n=5）,and the entrapment efficiency of lipoplexes was 89.94%.Conclusion： The formulation and the method can be used to prepare high quality cationic liposomes carrier,and to transfect cells effectively.Sephadex filtration is effective and simple and can be used to determine the entrapment efficiency of lipoplexes.

关 键 词：阳离子脂质体 质粒DNA 包封率 葡聚糖凝胶过滤法 体外转染 

分 类 号：R943.4[医药卫生—药剂学]