以乙肝核心抗原病毒样颗粒为新型载体蛋白制备流脑多糖结合疫苗  

Preparation of meningococcal polysaccharide conjugate vaccine using Hepatitis B core antigen virus-like particles as a novel carrier protein

作　　者：胥玲玲 李正军 苏志国[2] 杨延丽[2] 余蓉[1] 郑永祥 张松平[2] XU Lingling;LI Zhengjun;SU Zhiguo;YANG Yanli;YU Rong;ZHENG Yongxiang;ZHANG Songping(Key Laboratory of Drug Targeting and Drug Delivery System,Ministry of Education,West China School of Pharmacy,Sichuan University,Chengdu ,Sichuan,610041P.R.China;State Key Laboratory of Biochemical Engineering,Institute of Process Engineering,Chinese Academy of Sciences,Beijing,100190P.R.China;University of Chinese Academy of Sciences,Beijing,100049P.R.China)

机构地区：[1]四川大学华西药学院靶向药物及释药系统教育部重点实验室,四川成都610041 [2]中国科学院过程工程研究所生化工程国家重点实验室,北京100190 [3]中国科学院大学,北京100049

出　　处：《华西药学杂志》2018年第6期573-577,共5页West China Journal of Pharmaceutical Sciences

基　　金：国家自然科学基金资助项目(批准号:21336010、81773623)

摘　　要：目的以乙肝核心抗原(HBc)病毒样颗粒为载体,化学偶联脑膜炎奈瑟球菌荚膜多糖,制备高免疫原性的多糖结合疫苗。方法将双功能聚乙烯二醇对C群脑膜炎多糖衍生化后,与巯基化的乙肝核心抗原连接,采用凝胶过滤色谱纯化得到多糖结合疫苗。对结合疫苗的结构表征和多糖含量进行定量分析后,皮下免疫BALB/c小鼠,采用酶联免疫吸附法(ELISA)评价免疫效果。结果衍生化多糖与巯基化的HBc按质量比1:1投料,在4℃下反应48h,得到多糖蛋白比为0.69、游离多糖含量为14.66%的多糖结合疫苗,并且能够保持HBc完整的颗粒结构。免疫后,纯糖免疫组只能够产生有限的抗体滴度,而以HBc为载体的多糖结合疫苗能够诱导产生明显高于纯糖组的抗体滴度。结论采用HBc作为载体蛋白制备多糖结合疫苗,可以显著提高多糖的免疫原性。作为制备流脑多糖结合疫苗的新型载体蛋白,具有较好的研究价值和应用前景。OBJECTIVE To prepare highly immunogenic polysaccharide conjugate vaccine by chemically coupling with Neisseria meningitidis capsular polysaccharide using Hepatitis B core(HBc)antigen virus -like particles as a novel carrier protein.METHODS The meningitis group C meningitis polysaccharide(CPS)derivatized by heterobifunctional polyethylene glycol (PEG)was chemically linked to thiolated HBc.The obtained polysaccharide conjugate vaccine was then purified by gel filtration chromatography.After structural characterization and quantitative analysis,BALB/c mice were immunized subcutaneously and the immunogenicity of the conjugate vaccine was evaluated by enzyme -linked immunosorbent assay (ELISA).RESULTS By coupling the PEG derivatized CPS with thiolated HBc at a mass ratio of 1:1for 48h at 4℃,CPS -HBc conjugate vaccine with a polysaccharide protein ratio of 0.69and a free polysaccharide content of 14.66%,the structural integrity of the HBc VLPs was successfully synthesized well preserved.After the immunization,the plain CPS group could only produce limited antibody titers,while the antibody titer induced by CPS -HBc conjugate vaccine group was significantly higher than that of plain CPS group.CONCLUSION HBc can significantly improve the immunogenicity of polysaccharides ;therefore,the HBc can be a potential novel carrier protein with great research value and good application prospects.

关 键 词：流行性脑脊髓膜炎 C群脑膜炎多糖 乙肝核心抗原 病毒样颗粒 结合疫苗 载体蛋白 免疫活性 

分 类 号：R913[医药卫生—药学]