国产原材料在肠道病毒71型灭活疫苗(Vero细胞)生产中的替代研究

Substitution of domestic raw materials in the production of inactivated enterovirus 71 vaccine(Vero cell)

目的评估国产原材料(199培养基、DMEM培养基和球状微载体)替代进口原材料在肠道病毒71型(enterovirus 71,EV71)灭活疫苗(Vero细胞)规模化生产的可行性。方法在其他生产物料、生产设备、工艺流程和主要质控点等均不变的前提下,采用国产原材料进行3批次EV71灭活疫苗规模生产,对比分析进口原材料(变更前)和国产原材料(变更后)生产批次中间产品、成品的各项质量指标及产品稳定性。结果变更前后中间产品和成品的各项检测结果均符合质量标准要求,病毒收获液的抗原含量和病毒滴度平均值分别为6903 U/mL、8.7 lgCCID_(50)/mL和6171 U/mL、8.6 lgCCID_(50)/mL;原液的抗原含量、蛋白含量、比活、Triton X-100残留和纯度平均值分别为17605 U/mL、15μg/mL、1180 U/μg、7.0μg/mL、99.93%和15631 U/mL、14μg/mL、1111 U/μg、6.7μg/mL、100.00%;半成品的pH和铝含量平均值分别为6.8、0.18 mg/剂和6.9、0.18 mg/剂;成品的pH、体外相对效力、体内效力、渗透压摩尔浓度和铝含量平均值分别为6.9、388.0 U/剂、28.67 U、4.7 EU、333.7 mOsmol/kg、0.18 mg/剂和6.9、388.7 U/剂、25.00 U、6.2 EU、335.7 mOsmol/kg、0.18 mg/剂,均符合质量标准要求。变更前后产品长期稳定性和加速稳定性的结果均符合质量标准要求,且具有较好的一致性。结论国产原材料具有应用于EV71灭活疫苗(Vero细胞)规模生产的可行性。

Objective To evaluate the feasibility of replacing imported raw materials with domestic raw materials(199 medium,DMEM medium and microcarriers)in the large-scale production of inactivated enterovirus 71 vaccine(Vero cell).Methods On the premise that other production materials,production equipment,process flow and main quality control points were unchanged,three batches of EV71 inactivated vaccine were produced with domestic raw materials.The quality indexes and product stability of intermediate products and finished products produced with imported raw materials(before change)and domestic raw materials(after change)were compared.Results All the test results of intermediate products and finished products before and after change were meet the quality standards.The average antigen content and viral titer of viral harvests were 6903 U/mL,8.7 lgCCID_(50)/mL and 6171 U/mL,8.6 lgCCID_(50)/mL,respectively.The average values of antigen content,protein content,specific activity,Triton X-100 residue and purity of the bulk were 17605 U/mL,15μg/mL,1180 U/μg,7.0μg/mL,99.93%and 15631 U/mL,14μg/mL,1111 U/μg,6.7μg/mL,100.00%,respectively.The average values of pH and aluminum content of the final bulk were 6.8,0.18 mg/dose and 6.9,0.18 mg/dose,respectively.The average values of pH,relative in vitro potency,in vivo potency,osmolality and aluminum content of the final product were 6.9,388.0 U/dose,28.67 U,4.7 EU,333.7 mOsmol/kg,0.18 mg/dose and 6.9,388.7 U/dose,25.00 U,6.2 EU,335.7 mOsmol/kg,0.18 mg/dose,respectively.The results of long-term stability and accelerated stability of product before and after change meet the quality standards and have good consistency.Conclusion Domestic raw materials are feasible for large-scale production of EV71 inactivated vaccine(Vero cell).