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星点设计优化非病毒载体前阳离子脂质体的制备工艺 被引量:3

Application of star point design in optimization the formulation of non-viral gene delivery vector:the procationic liposomes
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摘要 目的采用星点设计优化前阳离子脂质体的制备工艺并考察前阳离子脂质体的有关性质。方法以薄膜分散膜挤压法制备前阳离子脂质体,以平均粒径、转染效率为指标,考察鱼精蛋白(Protamine)与质粒DNA的比例、脂质体骨架材料CHETA浓度、CHETA与质粒DNA的比例3个因素对制备条件的影响,将实验结果用数学方法处理并进行方程模型拟合,根据拟合方程及反应曲面图确定指标的最优值和各因素相对应的最佳取值范围。以LacZ为报告基因转染肝癌细胞HepG2,定量测定β-半乳糖苷酶活性和总蛋白含量,并计算转染效率。结果影响平均粒径、转染效率的3个主要因子的最佳取值分别为:Protamine/DNA(2.5:1);CHETA(45%);CHETA/DNA(4:1)。所制前阳离子脂质体形态近似于球体,平均粒径228.9±8 nm,多分散指数为0.122±0.02,Zeta电位为?25.08±2.5 mV,载基因前阳离子脂质体转染HepG2肝癌细胞转染效率为24.26±2.6 mU.mg-1。结论星点设计应用简便、预测性好,制备的前阳离子脂质体符合设计要求。 OBJECTIVE To optimize the preparation of proeationie liposomes by using star point design and to investigate the related characteristics. METHODS Blank procationic liposomes were prepared by film dispersion - fiheration method. The effects of Protamine/DNA ratio, CHETA molar percent and CHETA/DNA ratio on the mean particle size, and transfection efficiency were investigated. A second - order polynomial equation was fitted to the data and the resulting model was used to predict the response in the optimal region. Estimation of transfection efficiency was performed by galactosidase assay in HepG2 cells using LacZ as reporter gene. RESULTS All the investigated response variables were found to be highly dependent on the formulation variables. The optimized value of Protamine/DNA ratio, CHETA molar percent and CHETA/DNA ratio were(2.5 : 1), 45% and(4 : 1), respectively. The resulting proeationie liposomes had a regular spherical surface with an average size of 228.9 ± 8 nm (PDI of 0. 122 ± 0. 02), and a Zeta potential of - 25.08 ± 2.5 mV. The transfection efficiency of the delivery system, composed of procationic liposomes in HepG2 cells was 24.26 ± 2.6 mU· mg^ - 1 CONCLUSION Star point design is successfully used to optimize the preparation of procationic liposomes.
出处 《华西药学杂志》 CAS CSCD 北大核心 2007年第5期496-498,共3页 West China Journal of Pharmaceutical Sciences
关键词 星点设计 效应面 前阳离子脂质体 鱼精蛋白 转染效率 Star point design Response Surface Procationic liposomes Protamine Transfection efficiency
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