23价肺炎球菌多糖疫苗的规模化生产及质量控制

目的:扩大肺炎球菌发酵的培养规模,增加肺炎球菌多糖的收量,降低成品疫苗内毒素的含量,以提高肺炎球菌多糖疫苗的产能和质量;制备和标定企业用肺炎多糖参考品,及定性定量检测23价肺炎球菌多糖疫苗用的血清。方法:调整肺炎球菌培养参数及培养过程中糖、碱的补料方式、补料时间,放大肺炎球菌的发酵培养规模;在分段乙醇沉淀过程中添加速度和温度控制、调整乙醇浓度,在酚提过程中调控各种技术参数等,提高精制多糖的收量;控制加入生产过程中主要原辅材料的内毒素含量,并规范操作人员的行为习惯,降低肺炎球菌多糖疫苗成品的内毒素含量。标定自制多糖参考品以及用多糖免疫兔、鼠制备血清参考品。结果:肺炎球菌的发酵体积从500 L提高到1 000 L;精制多糖平均收量提高15%以上;内毒素平均含量控制在20 EU.mL-1以内;制备出23种型别的符合23价肺炎球菌多糖疫苗质量标准的多糖;获得3个型单抗和6个型的多抗。结论:23价肺炎球菌多糖疫苗的产量增加,疫苗质量得到了提升,自主化标准参考品研究进展迅速。

Scale production and quality control of 23-valent pneumococcal polysaccharide vaccine

Objective: To improve the productivity and the quality of pneumococcal polysaccharide vaccine for expanding the scale of pneumococcal fermentation,increasing the yield of polysaccharides and reducing the endotoxin content of vaccine;to prepare and calibrate the pneumococcal polysaccharide reference products for enterprises and the serum for qualitatively and quantitatively detecting the 23-valent pneumococcal polysaccharide vaccine.Methods: Pneumococcal amplification fermentation was expanded,and the culture parameters,the feeding mode,feeding time of sugar and alkali in the cultivation were adjusted.The yield of purified polysaccharide was increased by adding the speed control and the temperature control,adjusting the ethanol concentration in the subparagraph ethanol precipitation process,and controlling the various technical parameters in the phenol extraction process.The endotoxin content in pneumococcal polysaccharide vaccine was reduced by controlling the main raw materials used in the production process and standardizing the behavior of operators.Rabbits and rats were immunized by polysaccharide to obtain the standard serum.Results: Pneumococcal fermentation volume increased from 500 to 1 000 L.Average yield of purified polysaccharides was more than 15%.Average endotoxin content was less than 20 EU·mL-1.The polysaccharide that met the 23-valent polysaccharide pneumococcal polysaccharide vaccine quality standards had been prepared.Three monoclonal antibodies and 6 polyclonal antibodies had been prepared.Conclusion: The yield of 23-valent pneumococcal polysaccharide vaccine is increased,and the quality is improved.The progress in domestic standard reference develops rapidly.