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

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450 μg/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，Tm=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。

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

Objective  To investigate the antigen adsorption effect, stability and immunogenicity of recombinant human papillomavirus 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% effectious dose in mice were detected. Results  When the HPV stock solution mixed with 450 μg/ml aluminum hydroxide in histidine buffer system at pH6.5, the experimental vaccine had uniform appearance and the highest stability of antigen proteins, melting temperature=87.4 ℃. 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 was＞99.00%. Conclusion  The suitable conditions for  preparation of recombinant HPV vaccine are determined.