负载pVAX1-wapA的壳聚糖及季铵化壳聚糖纳米粒的制备研究  

Preparation of DNA-loaded chitosan nanoparticle vaccine

作　　者：向婧洁[1] 钟延强[1] 陆一鸣[1] 鲁莹[1] 

机构地区：[1]第二军医大学药学院,上海200433

出　　处：《药学实践杂志》2016年第1期19-23,40,共6页Journal of Pharmaceutical Practice

摘　　要：目的制备负载抗龋DNA疫苗pVAX1-wapA质粒的壳聚糖和季铵化壳聚糖纳米粒,优化其制备工艺,测定其细胞转染效率。方法以包封率和粒径为主要指标,单因素法考察载体浓度、pH值、N/P、TPP浓度等因素的影响,RealtimePCR检测细胞对质粒编码蛋白的转录表达水平以评价载质粒纳米粒的促转染作用。结果制得的载DNA疫苗纳米粒粒径均一,形态圆整。壳聚糖(CS)纳米粒粒径为(219.2±18.2)nm,Zeta电位为(24.7±3.5)mV,包封率为91.24%。季铵化壳聚糖(CSTM)纳米粒粒径为(222.5±15.6)nm,Zeta电位为(19.6±1.2)mV,包封率为87.66%。纳米粒可以促进pVAX1-wapA进入细胞,并成功被转录。结论制备的包载pVAX1-wapA的季铵化壳聚糖纳米粒可用于重组基因疫苗的运送。Objective To study and optimize the preparation condition of pVAXl-wapA-loaded nanoparticles and deter- mine the transfection efficiency. Methods The related effects of the crucial factors for the formation of nanoparticles： concen- tration of chitosan and TPP, pH value, N/P ratio were studied by single-factor experiment, with nanoparticles size and zeta potential as index. Cell transfection test was carried out to indicate that enhancement of cell transfection efficiency of nano-car- rier. Results Nanoparticles loaded DNA vaccine were nearly spherical shape with uniform particle size chitosan nanoparticle （CS） ,（219.2±18.2） nm;quaternary ammonium chitosan nanoparticles（CSTM）, （222.5±5.6） nm. Zeta potential of CS and CSTM was （24.7±3.5） mV, （19.6±1.2） mV and encapsulation efficiency was 91.24%, 87.66% ,respectively. CSTM nano- particle could enhance cellular uptake of pVAXl-wapA obviously. Conclusion CSTM nanoparticle was proved to be an efficient DNA vaccine delivery vector.

关 键 词：DNA疫苗 壳聚糖纳米粒 wapA蛋白 转染 季铵化壳聚糖 

分 类 号：Q78[生物学—分子生物学]