星点设计优化非病毒载体前阳离子脂质体的制备工艺
Application of star point design in optimization the formulation of non-viral gene delivery vector:the procationic liposomes
作者:刘中兵;宋庆国;
Author:
收稿日期: 年卷(期)页码:2007,(05):-496-498
期刊名称:华西药学杂志
Journal Name:WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字:星点设计;效应面;前阳离子脂质体;鱼精蛋白;转染效率
Key words:
基金项目:
中文摘要
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。
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