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聚乙烯亚胺/DNA复合物处方工艺优化及体外转染效率
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引用本文:李飞燕,周建平,吴向明,关丽叶,邓祖媛,谢 芳.聚乙烯亚胺/DNA复合物处方工艺优化及体外转染效率[J].中国药科大学学报(中文版),2010,41(1):40-44
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作者单位
李飞燕 中国药科大学药剂学教研室 
周建平 中国药科大学药剂学教研室 
吴向明 中国药科大学药剂学教研室 
关丽叶 中国药科大学药剂学教研室 
邓祖媛 中国药科大学药剂学教研室 
谢 芳 中国药科大学药剂学教研室 
基金项目:国家自然科学基金资助项目(No.30672548);江苏省“六大人才高峰”资助项目;国家“重大新药创制”科技重大专项资助项目(No.2009ZX09310-004)
中文摘要:目的:考察聚乙烯亚胺(PEI)相对分子质量、氮磷比(N/P比)、溶剂、离子强度等对PEI/DNA复合物形成、表面性质以及细胞转染效率的影响。方法:制备不同相对分子质量PEI/DNA复合物,通过凝胶电泳和紫外吸收检测确定PEI与DNA的复合能力(N/P比),测定不同溶剂、离子强度下的粒径和Zeta电位,考察复合物在HepG2细胞中的转染情况。结果:PEI与DNA的复合能力与PEI的相对分子质量呈正相关,不同溶剂、离子强度会影响复合物的表面性质,以磷酸盐缓冲液(PBS)为溶剂、PEI(25 kD)为载体、N/P比为12~15时,PEI/DNA复合物细胞转染效率明显优于质粒DNA,仅略低于阳性对照组。结论:经优化的PEI/DNA复合物可显著提高DNA在细胞中的转染效率。
中文关键词:聚乙烯亚胺  DNA复合物  工艺优化  细胞转染
 
Formula optimization and transfection efficiency of polyethyleneimine/DNA complexes
Abstract:Abstract Aim:To study the effects of molecule weight,the N/P ratio,solvent and ionic strength on the formation and surface properties of PEI/DNA complexes,and study its transfection efficiency.Methods:The N/P ratio of the prepared PEI/DNA complexes was optimized using gel electrophoresis and UV quantitative assay.The particle size and surface charge of the complexes were measured in different solvents and ionic strength.The transfection efficiency in HepG2 cells was observed.Results:There was a positive correlation between combination of PEI and DNA and molecule weight of PEI.In addition,the surface properties of PEI/DNA complexes were also influenced by the solvents and ionic strength.Comparable transfection efficiencies in HepG2 cells were observed for the Lipofectamine 2000/DNA and the prepared PEI(25 kD)/DNA complexes in PBS at the N/P ratio of 12-15,which was much higher than that of naked DNA.Conclusion:The optimized PEI/DNA complexes could effeciently transfect the cells in comparison to the positive control.
keywords:polyethyleneimine  DNA complexes  formula optimization  transfection efficiency in cells
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