质粒DNA构型分离和分析方法研究进展  

Research Progress of Plasmid DNA Conformation Separation and Analysis Methods

作　　者：王乐乐[1] 闻艳丽[1] 李兰英[1] 杨雪 李妍[1] 刘刚[1] WANG Le-le;WEN Yan-li;LI Lan-ying;YANG Xue;LI Yan;LIU Gang(Biometrology Laboratory,Shanghai Institute of Measurement and Testing Technology,Shanghai 201203,China)

机构地区：[1]上海市计量测试技术研究院生物计量实验室,上海201203

出　　处：《化学试剂》2021年第11期1509-1517,共9页Chemical Reagents

基　　金：上海市市场监督管理局科技项目(2019-12);上海市“科技创新行动计划”技术标准项目(18DZ2201700)。

摘　　要：作为基因传递的载体,质粒DNA已被广泛应用于DNA疫苗和基因治疗。质粒DNA存在不同的构型,其中超螺旋构型的质粒在细胞中的转染和表达效率最高。美国食品与药品监督管理局在2007年发布了质粒DNA疫苗的工艺指导原则,建议超螺旋构型的比例大于80%,因此建立高灵敏度和高分辨率的方法用于质粒DNA构型分离和分析具有重要意义。就近年来质粒DNA构型的分离和分析方法进行了综述,主要包括琼脂糖凝胶电泳法、液相色谱法和毛细管凝胶电泳法,并对未来质粒DNA构型分离和分析方法的发展提出了展望。As a gene delivery vector,plasmid DNA has been widely used in DNA vaccines and gene therapy.Plasmid DNA exists in different conformations,and the supercoiled plasmid has the highest transfection and expression efficiency in cells.The US Food and Drug Administration issued the guidance for plasmid DNA vaccines iN_(2)007,suggesting that the proportion of supercoiled conformation should be higher than 80%.Therefore,it is of great significance to establish a high-sensitivity and high-resolution method for plasmid DNA conformation separation and analysis.The separation and analysis methods of plasmid DNA conformation in recent years was reviewed,including agarose gel electrophoresis,liquid chromatography and capillary gel electrophoresis.The prospect for the future development of plasmid DNA conformation separation and analysis methods was also proposed.

关 键 词：质粒DNA 构型 琼脂糖凝胶电泳法 液相色谱法 毛细管凝胶电泳法 

分 类 号：O657.8[理学—分析化学]