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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 cationic liposomes encapsulating gene plasmids and investigate their properties, and determine the entrapment efficiency. Methods: DC-Chol and DOPE were used as materials to prepare cationic liposomes by membrane dispersion method. The liposomes were combined with gene plasmids expressing enhanced green fluorescent protein (GFP) and their transfection performance was investigated. The liposomes and lipids were measured by laser particle size analyzer The particle size and zeta potential of the complex were determined by gel filtration. The entrapment efficiency was determined by gel filtration method. Results: The average particle size of the prepared cationic liposomes and lipid complexes were 161.6 and 216.3 nm, respectively, and the Zeta potentials were +22.2 and +3.2 mV, respectively. The average particle size of the plasmid was 0.1925 ~ 3.85 μg.mL-1 Good linearity, high precision, no adsorption with dextran, column recovery was high, measured the encapsulation efficiency of the lipid complex was 89.94%. Conclusion: The cationic liposome carrier with good quality and effective transfection can be successfully prepared by using the prescription and the process. Sephadex GEL filtration method can accurately determine the entrapment efficiency. The method is rapid, simple and effective.