中检院发布《食品药品医疗器械检验机构仪器设备管理指南》.pdfVIP

432 中国医药生物技术 2014 年 12 月第 9 卷第 6 期 Chin Med Biotechnol, December 2014, Vol. 9, No. 6 Alkylated low molecular polyethylenimine as a gene delivery vector LIU Shan, HUANG Wei, JIN Ming-ji, QUAN Xiu-quan, GAO Zhong-gao 【Abstract 】 Objective Preliminary evaluation of the safety and efficiency of alkylated low molecular polyethylenimine as a gene delivery vector. Methods bPEI1.8K-C12 was synthesized with branch PEI (Mw 1800; bPEI1.8K) and 1-bromododecane, and structurally confirmed with 1H-NMR. Cytotoxicity and biocompatibility of bPEI1.8K-C12 were evaluated by MTT method and hemolysis assay, respectively. Size distribution and zeta potential of bPEI1.8K-C12/DNA polyplexes were determined, and cellular uptake of the DNA polyplexes was visualized with confocal laser scanning microscope (CLSM). DNA condensation ability of bPEI1.8K-C12 was studied by gel retardation assay, and in vitro gene delivery efficiency was investigated with luciferase and green fluorescent protein reporter gene, respectively. Results bPEI1.8K-C12 was successfully synthesized after confirmation by 1H-NMR. Cytotoxicity of bPEI1.8K-C12 on MCF-7 cells was as low as bPEI1.8K, and systematic delivery of high dose bPEI1.8K-C12 may cause hemolysis in vivo. The average size of bPEI1.8K-C12/DNA polyplexes was bigger than that of bPEI1.8K/DNA polyplexes at the same weight ratio, and the zeta potential of bPEI -C /DNA polyplexes was higher than that of bPEI /DNA polyplexes at the same weight ratio. After alkylation, the DNA 1.8K 12 1.8K condensation ability of bPEI1.8K-C12 was reduced, but the cellular uptake of bPEI1.8K-C12/DNA polyplexes was greatly enhanced. In vitro gene transfection efficiency of bPEI1.8K-C12/DNA was significantly higher than that of bPEI1.8K, and was comparable