空肠弯曲菌外膜蛋白亚单位脂质体佐剂疫苗的研制

Preparation of Subunit Vaccine with Liposomal Adjuvant for Campylobacter Jejuni

目的探索用脂质体包裹空肠弯曲菌(Campylobacterjejuni,CJ)28-31KD(即PEB1、PEB2、PEB3、PEB4)外膜蛋白制备安全、稳定、有效的CJ亚单位脂质体佐剂疫苗。方法采用薄膜-超声-冻融法制备阴离子脂质体(AL)、阳离子脂质体(CL)CJ28-31KD外膜蛋白亚单位佐剂疫苗,通过电子显微镜观察脂质体疫苗微粒的形态、测其粒径及分布,进行稳定性检测、无菌试验、热原质试验、疫苗的不良反应观察及包裹率检测等疫苗质量鉴定。结果1疫苗都为均匀乳白色混悬液,无分层和沉淀现象,阴离子脂质体疫苗颗粒形态呈球形,平均直径250nm,分布均匀,阳离子脂质体疫苗颗粒大,不规则;2两种疫苗的无菌试验及热原质试验均合格,不良反应观察结果均无明显不良反应;3阴、阳离子脂质体疫苗冻融前包裹率分别为32.1%、22.5%,冻融后分别是38.3%、28.4%。结论采用薄膜-超声-冻融法制备CJ亚单位佐剂疫苗是可行的;制备的疫苗是安全、稳定的;冻融法可以提高脂质体疫苗包裹率。

Objective： To explore the development of the outer membrane protein （28KD-31KD） with liposomes and prepare a safe , stable and effective subunit vaccine of campylobacter jejuni （CJ）. Methods.. The two liposomal subunit vaccines were prepared with the thin film diffusion-sonication-frozen melted method. The shape, diameter and distribution of the microparticulate of liposomal vaccines were observed by electronic microscope. The stability, Sterile test , Pyrogen test, encapsulation ratio and the side effects of the vaccines were identified. Result： The two kinds of vaccines were well-distributed milk-white liquid and not layered. Particles of anionic liposomal vaccine were round and its mean diameter is 250nm. Particles of cationic liposomal vaccine were oval-shaped and were bigger and more irregular than anionic liposomal vaccine. ①Pyrogen test,sterile test and the side effects of the two vaccines were qualified.② Before freezing,the encapsulation ratio of anionic, cationic liposomal vaccine were 32. 1% and 22. 5% respectively, but they were 38. 3% and 28. 4% after freezng. Conclusion： The thin film diffusion-sonication-frozen melted method can be used to prepare subunit vaccine of CJ. Both vaccines developed are safe and stable. Encapsulation ratio of the vaccines could be enhanced by frozen melted method.