犬细小病毒悬浮培养工艺研究
Study on suspension culture technology of canine parvovirus
李凤艳 1陈生雷 1刘艳霞 1刘梦成 1姜斌 1舒秀伟1
作者信息
- 1. 辽宁益康生物股份有限公司,辽宁辽阳 111000
- 折叠
摘要
研究旨在证明犬细小病毒(CPV)悬浮培养的可能性,提高CPVP6株抗原的病毒含量,降低转瓶生产不同批次间疫苗质量差异,生产质量稳定的疫苗产品.试验利用大孔的纤维编织物BioNOCⅡ型微载体所提供的巨大表面积,实现Vero细胞高密度培养,建立了 Tide-cell生物反应器大规模培养CPV P6株抗原制备工艺.结果显示:生物反应器Tide-cell载体罐内接入1.0×1010个F81种子细胞,在优化的参数条件下培养,用RPMI 1640培养液经过96h培养,细胞总数为1.0×1011个;在葡萄糖消耗量约为5 g/(L·h)时,按照病毒感染复数(MOI)=0.02将生产种毒CPVP6株接入Tide-cell微载体细胞培养瓶内,补加含8％血清的RPMI 1640病毒维持液至50 L,调整pH值为7.2～7.3,培养48h收获,病毒含量至少为107.43 TCID50/mL.
Abstract
The aim of the study was to demonstrate the possibility of suspension culture of CPV,improve the viral content of CPV P6 strain antigen,reduce the quality difference of vaccine between different batches produced by rotating bottle,and produce vaccine products with stable quality.The high density culture of Vero cells was realized by utilizing the huge surface area provided by BioNOC type Ⅱ microcarrier of fiber braided fabric with large pores,and the preparation process of CPV P6 strain antigen was established in a Tide-cell bioreactor.The results show that 1.0 × 1010 F81 seed cells were added into the bioreactor Tide-cell carrier tank and cultured under optimized parameter conditions.After 96 h culture in RPMI 1640 medium,the total number of cells was 1.0 × 1011 and glucose consumption was about 5 g/(L·h).The production seed virus CPV P6 strain was added into the Tidecell microcarrier cell culture vial according to the MOI=0.02,and the RPMI 1640 virus maintenance solution containing 8％serum was added to 50 L,the pH value was adjusted to 7.2～7.3,and the culture was harvested for 48 h.The virus content should be at least 107.43 TCID50/mL.
关键词
生物反应器/犬细小病毒/CPVP6株/微载体/疫苗Key words
Bioreactor/Canine parvovirus/CPV P6 strain/Microcarrier/Vaccine引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据