幽门螺杆菌尿素酶B亚单位W/O/W型复乳剂的研制

Preparation of W/O/W type multiple emulsions containing Helicobacter pylori urease B subunit

目的 制备重组幽门螺杆菌尿素酶B亚单位(rUreB)复乳剂,研究其物理性质及免疫效率。方法 设计处方采用二步法制备复乳,显微镜观察乳滴结构及粒径分布,以电导法测定包封率,以经典法、离心法及电导法观察复乳的稳定性,免疫印迹观察抗原制备成复乳剂后免疫反应性的变化,以动物实验观察rUreB复乳的免疫效率。结果 制备的rUreB复乳性质稳定、包封率98.3％、粒径大小主要在2～10μm,免疫印迹及小鼠口服免疫后特异性抗体测定结果表明,该复乳具有良好的免疫原性,剂量小、免疫效率高。结论 将幽门螺杆菌尿素酶B亚单位制备成复乳剂可得到高效、廉价、使用方便的疫苗,复乳是一个很有潜力的黏膜疫苗传送系统。

Objective To prepare W/O/W type multiple emulsion(ME)containing helicobacter pylori urease B subunit and study on it's physical properties and immune efficiency. Methods Multiple emulsion were prepared by two-stage emulsification method. The structure of emulsion and distribution of particle size were observed by microscope. Drug entrapment efficiency was measured through conductance rate. The stability of emulsion was studied by classical method, centrifugation method and conductance rate method. The antigenic integrity was evaluated by western blotting. The immune efficiency was evaluated by immunization in Balb/c mice. Results We prepared stable W/O/W emulsion with a particle size of 2-10μm and an entrapment efficiency of 98.3%. The results of immunoblotting and immunization showed that multiple emulsion containing rUreB markedly enhanced immunization efficiency. Conclusion The immune response of ME containing rUreB is stronger than that of rUreB solution. Multiple emulsion is a potential delivery system of mucosa vaccine.