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目的通过对发热伴血小板减少综合征布尼亚病毒(简称“新布尼亚病毒”)进行规模化生产培养工艺研究,为新布尼亚病毒规模化生产提供有力支持。方法采用细胞工厂培养Vero细胞,待其长成致密单层后,取工作种子批新布尼亚病毒毒种接种细胞,采用连续收获或细胞病变充分时收获培养液上清的方法收获病毒,并以病毒滴度、抗原含量作为评价指标选择基础培养基、培养基pH、人血白蛋白添加浓度、接种细胞日龄、接种病毒MOI以及病毒培养温度。结果按0.01~0.001 MOI接种3~4日龄Vero细胞,病毒培养液选择含0.3%人血白蛋白pH 7.6~7.8的DMEM溶液,35℃培养7 d收获,病毒收获液病毒滴度7.87 LgCCID50/mL、抗原含量170.1μg/mL。结论初步建立了新布尼亚病毒规模化生产培养工艺,为后续工业化生产提供了数据支持。
Objective To study the large-scale production and culture of fever virus with thrombocytopenia syndrome Bunyavirus (hereinafter referred to as “new Bunyan virus”), and provide strong support for the mass production of new Bunyan virus. Methods Vero cells were cultured in a cell factory. After the cells grew into a dense monolayer, the working seed seeds were inoculated with new Bunyavirus and the virus was harvested by harvesting the supernatant of the culture supernatant with continuous harvest or cytopathic effect The virus titer and antigen content were used as the evaluation indexes to select the basic medium, the medium pH, the concentration of human serum albumin, the days of inoculation, the MOI of virus inoculation and the temperature of virus culture. Results Vero cells of 3 to 4 days old were inoculated with 0.01 ~ 0.001 MOI. The DMEM solution containing 0.3% human serum albumin (pH 7.6 ~ 7.8) was selected in the virus culture medium and harvested at 35 ℃ for 7 days. The virus titer was 7.87 LgCCID50 / mL, antigen content 170.1μg / mL. Conclusion The scale-up culture process of new Bunyan virus was initially established, which provided the data support for the follow-up industrialized production.