甲型肝炎灭活疫苗(L-A-1减毒株)的制备及其免疫原性  

Preparation and immunogenicity of inactivated hepatitis A vaccine(attenuated L-A-1 strain)

作　　者：刘令九[1] 岳立广[1] 徐艳玲[1] 惠琦[1] 徐晓霞[1] 侯丽娟[1] 李淑焱[1] 李勇[1] 夏青娟[1] 赵小琳[1] 

机构地区：[1]长春生物制品研究所有限责任公司,吉林长春130012

出　　处：《中国生物制品学杂志》2015年第1期6-9,共4页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划项目(20100941)

摘　　要：目的用甲型肝炎病毒（hepatitis A virus,HAV）L-A-1减毒株制备甲型肝炎灭活疫苗,并检测其免疫原性。方法用细胞工厂培养人胚肺二倍体细胞（2BS株）,感染L-A-1减毒株增殖病毒,收获含病毒的细胞,经裂解、聚乙二醇（polyethylene glycol,PEG）6000沉淀、三氯甲烷抽提、凝胶过滤层析纯化后,甲醛灭活,氢氧化铝佐剂吸附,制备3批试验疫苗。按照《中国药典》三部（2010版）中甲肝灭活疫苗标准进行各项检定,并进行小鼠效力试验,计算半数有效稀释倍数;将疫苗于37℃存放5、10、15、20、25和30 d,考察疫苗的加速稳定性。结果甲型肝炎病毒经提取纯化后,抗原回收率达80%以上,杂蛋白去除率达95%以上;制备的3批试验疫苗各项检定结果均符合《中国药典》三部（2010版）要求;小鼠效力试验结果显示,半数有效稀释倍数分别为进口疫苗和国产疫苗的1.25～1.40和2.02～2.26倍;37℃保存30 d,疫苗的体外相对效力仍在合格范围内。结论制备了纯度较高的甲型肝炎灭活疫苗,其在小鼠体内具有良好的免疫原性,为进一步规模化生产甲型肝炎灭活疫苗奠定了基础。Objective To prepare inactivated hepatitis A vaccine with attenuated L-A-1 strain and determine its immunogenicity. Methods Human embryo lung diploid cells （2BS strain） were cultured with cell factory and infected with attenuated L-A-1 strain. The cells containing propagated virus were harvested, lysed, precipitated with polyethylene glycol （PEG） 6000, extracted with chloroform, purified by gel filtration chromatography, inactivated with formaldehyde and adsorbed with aluminium hydroxide. Three batches of experimental vaccine were prepared and subjected to overall control tests according to the requirements in Chinese Pharmacopoeia （Volume Ⅲ, 2010 edition）. Potency test in mice was performed, based on which the median effective dilution was calculated. The vaccine was stored at 37 ℃ for 5, 10, 15, 20, 25 and 30 d to evaluate the accelerated stability. Results The antigen recovery rate of hepatitis A virus after extraction and purification was more than 80%, while the removal rate of foreign protein reached more than 95%. All the quality indexes of three batches of experimental vaccines met the requirements in Chinese Pharmacopoeia （Volume Ⅲ, 2010 edition）. The potency test in mice showed that the median effective dilutions were 1.25 ～ 1.40 and 2. 02 ～2. 26 times of those of imported and domestic vaccines respectively. The relative potency in vitro of vaccine after storage at 37 ℃ for 30 d was still qualified. Conclusion Inactivated hepatitis A vaccine with high purity was prepared, which showed good immunogenicity in mice. It laid a foundation of further large-scale production of inactivated hepatitis A vaccine.

关 键 词：L-A-1减毒株 甲型肝炎灭活疫苗 免疫原性 

分 类 号：R373.21[医药卫生—病原生物学] R392-33[医药卫生—基础医学]