麻疹、腮腺炎、风疹、水痘联合减毒活疫苗的生产工艺研究

目的 建立麻疹、腮腺炎、风疹、水痘联合减毒活疫苗(combined live attenuated measles,mumps,rubella and varicella vaccine,MMRV)的生产工艺.方法 根据现有疫苗原液生产工艺,将麻疹病毒沪-191纯化株、腮腺炎病毒S79株、风疹病毒BRD-Ⅱ株和水痘-带状疱疹病毒Oka株在原代鸡胚成纤维细胞或人二倍体细胞MRC-5株中制备高滴度病毒原液,并超低温保存.筛选无明胶冻干稳定剂配方.按国外已上市同类产品的病毒配比,研究MMRV中4种病毒的原液配制滴度及成品配制比例,建立最佳冻干工艺.结果 用筛选出的适于MMRV的无明胶冻干稳定剂配方进行试验,确定病毒的原液配制滴度为,麻疹4.6 lg半数细胞培养感染量(50% cell culture infective dose,CCID50)/ml、腮腺炎5.8 lgCCID50/ml、风疹4.3 lgCCID50/ml、水痘4.8 lg噬斑形成单位(plaque forming unit,PFU)/ml.使成品中腮腺炎病毒滴度至少达到麻疹和风疹病毒的10倍,水痘病毒滴度高于现有单价水痘疫苗.连续制备3批MMRV,经检测平均病毒滴度为,麻疹4.5 lgCCID50/ml、腮腺炎5.1 lgCCID50/ml、风疹4.3 lgCCID50/ml、水痘4.6 lgPFU/ml;平均水分为1.2%.其他项目检定均合格.结论 建立了MMRV的生产工艺,可以稳定生产出达到国外同类产品质量标准并符合我国4种单价减毒活疫苗国家标准的产品.

Research on production process of combined live attenuated measles,mumps, rubella and varicella vaccine

Objective  To establish the production process of combined live attenuated measles, mumps, rubella and varicella vaccine (MMRV). Methods The high titer vaccine virus bulks were prepared by culturing purified measles virus Hu-191 strain, mumps virus S79 strain, rubella virus BRD-II strain and varicella-zoster virus Oka strain in primary chicken embryo fibroblast cell or human diploid cell MRC-5 strain, and stored in ultra low temperature freezer. The gelatin-free freeze-drying stabilizer formulation was screened. According to the same class overseas products, the titers and formulations of 4 virus bulks were researched and the best freeze-drying process was established. Results   Using the gelatin-free freeze-drying stabilizer formulation suitable for MMRV, the virus titers in vaccine bulks were 4.6 lg 50% cell culture infective dose (CCID₅₀)/ml for measles, 5.8 lgCCID₅₀/ml for mumps, 4.3 lgCCID₅₀/ml for rubella, and 4.8 lg plaque forming unit (PFU) /ml for varicella. The mumps virus titer in final product was at least 10 times of measles and rubella virus titers. The titer of varicella virus was higher than that of monovalent varicella vaccine. The average virus titers of 3 continuous batches of MMRV were 4.5 lgCCID₅₀/ml for measles, 5.1 lgCCID₅₀/ml for mumps, 4.3 lgCCID₅₀/ml for rubella, and 4.6 lgPFU/ml for varicella. The average moisture was 1.2%. All other items tested were qualified. Conclusions The MMRV production process is developed, which can stably produce the products meeting the quality standards of similar foreign products and the national standards of 4 monovalent live attenuated vaccines in China.