生物反应器细胞培养制备人用狂犬病疫苗 |
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引用本文: | 查力,高军,侯剑英,白珠穆,袁德明,杨俊伟,李旭,李庆岸,陈焕玉,孙非非,赵红梅,周德水. 生物反应器细胞培养制备人用狂犬病疫苗[J]. 中国生物制品学杂志, 2006, 19(3): 288-290 |
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作者姓名: | 查力 高军 侯剑英 白珠穆 袁德明 杨俊伟 李旭 李庆岸 陈焕玉 孙非非 赵红梅 周德水 |
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作者单位: | 辽宁成大生物技术有限公司 (沈阳110026)(查力,侯剑英,袁德明,李旭,陈焕玉,赵红梅,周德水),辽宁省生物医学工程研究院有限公司 (沈阳110026)(高军,白珠穆,杨俊伟,李庆岸,孙非非) |
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基金项目: | 辽宁省海外学子创业基金 |
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摘 要: | 目的应用生物反应器培养细胞和病毒,大规模生产人用狂犬病疫苗。方法以巴斯德PV2061为毒种,以143代以内Vero细胞为培养基质,应用生物反应器,每升投放25g微载体,灌流式细胞培养,连续收获病毒液,经浓缩、灭活、纯化,制成Vero细胞狂犬病疫苗。结果细胞培养密度达1.2×107~1.5×107个/ml,病毒感染后可连续收获18~22d,病毒最高滴度8.5LogLD50/ml,平均滴度7.6LogLD50/ml。经柱层析纯化,杂蛋白去除率达99.95%以上,总蛋白含量≤80μg/g,DNA含量≤10pg/0.5ml,GP含量3.5~4.5IU/0.5ml,效力≥4.5IU/0.5ml。结论应用生物反应器细胞培养,可以大规模生产优质Vero细胞人用狂犬病疫苗。
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关 键 词: | 生物反应器 微载体 Vero细胞 狂犬病疫苗 |
收稿时间: | 2006-01-12 |
修稿时间: | 2006-01-12 |
Preparation of Rabies Vaccine for Human Use by Cell Culture in Bioreactor |
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Abstract: | Objective To prepare rabies vaccine for human use in a large scale by cell culture in bioreactor.Methods Prepare rabies vaccine by perfused culture of Vero cells within 143 passages inoculated with PV2061 strain in bioreactor containing 25 g/L of microcarrier.The virus bulk was continuously harvested,then concentrated,inactivated and purified.Results The density of cultured cells reached 1.2×107~1.5×107 cells/ml.The virus bulk could be continuously harvested for 18~22 d after inoculation and reached a highest titer of 8.50 LogLD 50/ml and a mean titer of 7.2~7.6 LogLD 50/ml.After purification by column chromatography,the removal rate of foreign protein reached more than 99.95%.The total protein,DNA and GP contents of final bulk were not more than 80 μg,not more than 10 pg and 3.5~4.5 IU respectively,and its potency was not less than 4.5 IU/0.5 ml.Conclusion The cell culture in bioreactor could be used for the large-scale production of rabies vaccine for human use. |
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Keywords: | Bioreactor Microcarrier Vero cells Rabies vaccine |
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