高等学校化学学报

高等学校化学学报 ›› 2003, Vol. 24 ›› Issue (11): 1993.

• 论文 • 上一篇    下一篇

PEG化壳聚糖/DNA自组装复合物的制备、表征和体外Hela细胞转染研究

魏晓红1, 梁文权1, 潘远江2   

  1. 1. 浙江大学药学院, 杭州 310031;
    2. 浙江大学理学院化学系, 杭州 310012
  • 收稿日期:2002-12-03 出版日期:2003-11-24 发布日期:2003-11-24
  • 通讯作者: 梁文权(1945年出生),男,教授,博士生导师,从事药物的控制释放和生物技术药物的靶向给药研究.E-mail:wqliang@cmm.zju.edu.cn E-mail:wqliang@cmm.zju.edu.cn
  • 基金资助:

    国家自然科学基金(批准号:39970879)资助

Preparation and Characterization of PEGylated Chitosan/DNA Self-assemble Complex and the Research on Transfection on Hela CelLIn Vitro

WEI Xiao-Hong1, LIANG Wen-Quan1, PAN Yuan-Jiang2   

  1. 1. College of Pharmacy, Zhejiang University, Hangzhou 310031, China; Department of Chemistry, College of Science, Hangzhou 310012, China
  • Received:2002-12-03 Online:2003-11-24 Published:2003-11-24

PDF (PC)

被引次数

50

摘要： 通过有机合成和高分子聚合等方法将亲水性的聚乙二醇接枝到壳聚糖的氨基侧链上,得到了改性的壳聚糖-聚乙二醇接枝共聚物,应用现代波谱等技术对中间产物和最终产物进行了表征.采用绿色荧光蛋白基因质粒pEGFP-N1为DNA模型,在溶液中通过自动(静电)吸附得到PEG化的壳聚糖/DNA自组装复合物.初步研究了该自组装复合物对Hela细胞的体外转染效率.结果表明,活化的聚乙二醇被成功地接枝到壳聚糖上,使不溶于水的壳聚糖改性为水溶性的PEG化的壳聚糖.PEG化壳聚糖/DNA自组装复合物在Hela细胞体外转染率达到81%.因此,PEG化的壳聚糖有可能成为基因转染的非病毒载体.

关键词: 聚乙二醇活化物, 壳聚糖, 自组装复合物, 非病毒类载体, 体外转染

Abstract: Chitosan, as a non-viral delivery system for gene therapy, has been increasingly proposed because of its biodegradable, cationic, non-toxic, good biocompatibility.As hydrophobic polymers are usually taken up by reticuloendothelial-system(RES) and have a short residence time in blood, hydrophilic, flexibleand non-ionic polymer, methoxypolyethyleneglycol(mPEG) was grafted to the amino group of chitosan to modify the chitosan′s property in this paper.mPEG-Chitosan is obtained through three steps of esterification reactions.The structures of the activated mPEGand mPEG-Chitosan were confirmed with 1HNMR, 13CNMR and Fourier transform infrared(FTIR) spectrum.The solubility of PEGy lated chitosan in water is 53.6mg/mL, which becomes dissolved in water while chitosan is undissolvable.The percentage of PEGgrafted onto the amino group of chitosan was 16.71%(molar fraction).Plasmid pEGFP-N1was chosen as model DNA.The PEGy lated chitosan/DNA complex was prepared by using an auto-coacervation process.The transfection efficiency of the PEGy lated chitosan/DNA complex in Hela cellSIn vitro was 81% analyzed by flow cytometer.Thus, The PEGy lated chitosan could be a candidate of effective non-viral vectors for gene delivery system.

Key words: PEGylated compound, Chitosan, Self-assemble complex, Non-viral vector, Transfection in vitro

中图分类号: 

TrendMD: