重组四价人乳头瘤病毒16/18/52/58型病毒样颗粒疫苗制剂工艺的优化  

Optimization of preparation process of recombinant human papillomavirus 16/18/52/58 virus-like particle vaccine

作　　者：王蓓 王文伟 仝光杰 胡海涛 楼觉人 Wang Bei;Wang Wenwei;Tong Guangjie;Hu Haitao;Lou Jueren(Research and Development Department,Shanghai Wellvax Co.,Ltd.,Shanghai 200051,China;No.2 Laboratory,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 200051,China)

机构地区：[1]上海至成生物科技有限公司研发部,200051 [2]上海生物制品研究所有限责任公司第二研究室,200051

出　　处：《国际生物制品学杂志》2021年第6期323-328,共6页International Journal of Biologicals

基　　金：上海市科学技术委员会科研计划项目(19431907800)。

摘　　要：目的考察不同制剂工艺流程下制备的重组四价人乳头瘤病毒(human papillomavirus,HPV)16/18/52/58型病毒样颗粒疫苗的抗原吸附效果、稳定性及免疫原性,根据实验结果筛选出最合适的制剂配方及工艺。方法通过改变重组四价HPV疫苗的缓冲体系,以及氢氧化铝佐剂的含量、混合工艺、吸附方式与吸附条件,配制不同的实验疫苗,观察其外观,并分别进行蛋白质稳定性、抗原吸附率和小鼠体内半数有效剂量的检测。结果重组四价HPV疫苗原液与450μg/ml氢氧化铝佐剂在pH6.5的组氨酸缓冲体系下混合后的实验疫苗,外观均一性良好,抗原蛋白稳定性最高,熔解温度为87.4℃,小鼠体内免疫原性较好;搅动吸附条件下的4型别HPV抗原吸附率明显高于静止吸附;铝佐剂和抗原蛋白的吸附与混合先后顺序,不同吸附比例、温度和时间对疫苗外观无显著影响,抗原吸附率均大于99.00%。结论确定了重组四价HPV16/18/52/58型病毒样颗粒疫苗合适的制剂工艺条件。Objective To investigate the antigen adsorption effect,stability and immunogenicity of recombinant human papillomavirus(HPV)16/18/52/58 vaccine prepared by different preparation processes,and to select optimal formulation and process.Methods Different experimental vaccines were prepared by changing the buffer system,aluminum adjuvant content,mixing process,adsorption mode and conditions of recombinant HPV vaccine.The appearance of the vaccine was observed,and the antigen adsorption rate,protein stability and 50%effective dose in mice were detected.Results When HPV stock solution was mixed with 450μg/ml aluminum hydroxide in pH6.5 histidine buffer system,the experimental vaccine had uniform appearance,the highest stability of antigen proteins(melting temperature=87.4℃),and good immunogenicity in mice.The adsorption rate of HPV antigen under stirring adsorption was much higher than that under static condition.The sequence of adsorption and mixing of aluminum adjuvant and antigen protein,as well as different adsorption ratios,temperatures and time,had no significant effect on the appearance of the vaccine.The antigen adsorption rate were>99.00%.Conclusion The suitable conditions for preparation of recombinant HPV 16/18/52/58 vaccine are determined.

关 键 词：人乳头瘤病毒 佐剂 免疫 抗原吸附率 

分 类 号：R373[医药卫生—病原生物学]