大熊猫犬瘟热病毒（CDV-PA-1421株）微载体悬浮培养工艺研究

Study on Giant Panda Canine Distemper Virus (CDV-PA-1421 Strain) Microcarrier Suspension Culture

关键词:  大熊猫犬瘟热  微载体悬浮培养  生物反应器

Key words:  Giant Panda Canine Distemper Virus  Microcarrier Suspension Culture  Bioreactor

摘要:

根据正交试验对大熊猫犬瘟热病毒(CDV-PA-1421株)在生物反应器中的培养工艺进行优化，确定关键技术参数。结果表明，大熊猫犬瘟热病毒悬浮培养最佳组合条件为：Vero细胞长成单层后，接种到生物反应器，细胞接种浓度为0.2×10⁶/mL~0.4×10⁶/mL，微载体浓度为8.0 g/L，加入MSVP培养基至工作体积。按0.01～0.1 感染复数(MOI)接种病毒，在37 ℃ pH7.2条件下培养96~120 h，细胞病变(CPE)达70%～80%时收获病毒上清液，即可获得高滴度病毒液。3批培养结果显示病毒培养工艺稳定，收获液病毒含量均在10^7.0～10^7.5 TCID₅₀/0.1mL之间。

Abstract:

The orthogonal design was used to optimize the culture conditions of giant panda canine distemper virus (CDV-PA-1421 strain) in bioreactor, and key technical parameters were determined. Results showed that the best combined conditions for suspension culture of giant panda canine distemper virus were: Vero cells were grown into monolayers and inoculated to the bioreactor at a concentration of 0.2×10⁶/mL~0.4×10⁶/mL and a microcarrier concentration of 8.0 g/L. MSVP culture medium was added to the working volume. The virus was inoculated at 0.01~0.1 MOI and cultured at 37 ℃, pH 7.2 for 96~120 hours. When the CPE was up to 70%~80%, the virus supernatant was harvested to obtain the high-titer virus venom. The results of three batches of culture showed that the virus culture process was stable, and the virus content in the harvest solution was in the range of 10^7.0～10^7.5 TCID₅₀/0.1mL.