摘要
目的建立利用生物反应器制备Vero细胞乙型脑炎纯化疫苗的新工艺。方法以Vero细胞作为乙型脑炎病毒增殖的细胞基质,使用微载体Cytodex-Ⅰ在15L生物反应器中进行高密度培养,采用2.5～4.5g/L的载体浓度培养乙型脑炎病毒,制备3批纯化乙型脑炎疫苗并进行检定。结果随着微载体浓度的增加,细胞密度升高。采用2.5～4.5g/L微载体培养的病毒收获液的平均滴度为7.38～7.56lgPFU/ml,收获量最高可达到12～15个有效罐体积。制备的3批疫苗各项质量指标均符合《中国药典》三部(2005版)相关要求。结论已建立了15L生物反应器制备Vero细胞乙型脑炎纯化疫苗的新工艺,为进一步放大生产规模奠定了基础。
Objective To develop a new procedure for preparation of purified Japanese encephalitis （JE） vaccine with Vero cells by using bioreactor. Methods JE virus was propagated in Vero cells on mierocarrier Cytodex- Ⅰ at a concentration of 2. 5- 4. 5 g / L in a 15 L bioreactor for high density culture, and the harvested virus liquid was prepared into purified JE vaccine. Three batches of vaccine were prepared and subjected to control tests. Results The cell density increased with the increasing concentration of microearriers, The titer of harvested virus liquid cultured on microcarrlers was 7. 38 - 7. 56 IgPFU / ml, and the maximum yield reached 12-15 working volumes. The quality indexes of 3 batches of prepared vaccine met the relevant requirements in Chinese Pharmacopoeia （Volume Ⅲ, 2005 edition）. Conclusion A new procedure for preparation of purified JE vaccine with Veto cells by using 15 L bioreactor was developed, which laid a foundation of large-scale production of the vaccine.
出处
《中国生物制品学杂志》
CAS
CSCD
2009年第6期591-592,595,共3页
Chinese Journal of Biologicals
基金
甘肃省科技攻关计划项目(2GS063-A43-011)
