摘要
目的:制备LE-pGJA-P/VAX黏膜递呈系统,并对其理化和生物学性质进行检测。方法:利用中试生产的pGJA-P/VAX质粒通过与脂质体LE溶液直接混合制备LE-pGJA-P/VAX复合物,通过琼脂糖凝胶电泳阻滞实验和激光粒度仪检测该复合物的理化性质,并将其体外转染CHO细胞检测是否能体外表达目的蛋白。结果:制备的LE-pGJA-P/VAX复合物平均粒径为50nm,ZETA电位为14.5mV,体外转染CHO细胞48h后,经细胞免疫荧光染色,转染组可见细胞浆红色荧光染色,提示可有效表达质粒编码的PAc蛋白,而裸质粒转染组则未见明显红色荧光。结论:LE-pGJA-P/VAX复合物表面电荷均匀,粒径小,且粒径分布范围窄,符合纳米粒子特征,可在体外表达编码蛋白,提示该递呈系统可用于防龋DNA疫苗的黏膜递呈,为后续增强防龋DNA疫苗经鼻腔黏膜途径投递的免疫效能奠定基础。
Objective： To prepare a mucosal delivery system of LE-pGJA-P/VAX and study the physicochemical and biological characteristics. Methods： Anti-caries DNA vaccine pGJA-P/VAX were coated with liposomes emulsion （LE）, forming LE-pGJA-P/VAX. Agarose gel electrophoresis and a laser particles analyzer were used for showing the physicochemical characteristics. CHO cells were transfected with this complex in vitro to evaluate its biological characteristics. Results： The agarose gel electrophoresis showed most of the DNA plasmid in the preparation system was encapsulated in the complex. The mean particle size of the complex was 50nm, ZETA potential was 14.5inV. Immunofluorescence demonstrated the expressing of encoded PAc protein of LE-pGJA-P/VAX transfected cells, while naked plasmid transfected cells showed no fluorescence. Conclusion： The size distrubution of the LE-pGJA-P/VAX complex was narrow and surface charge was well-distrubuted. The mucosal delivery system of LE-pGJA-P/VAX was proved to be nanoparticles and be able to express encoded protein in vitro.
出处
《口腔医学研究》
CAS
CSCD
2012年第3期218-219,共2页
Journal of Oral Science Research
基金
国家自然基金资助项目(编号:81070822)
武汉市发展引导类计划资助项目(编号:201060938367)
关键词
防龋DNA疫苗
脂质体
黏膜
递呈系统
Anti-caries DNA vaccine Liposomes emulsion（LE） Mucosa Delivery system
