慢病毒载体大规模生产技术的探究  

Research on Large Scale Production Technology of Lentiviral Vector

作　　者：贾裕 金威洋 刘阳[1,2,3] 陈阳 贾旭 孙杰[1,2,3,4] 佟莹莹 王华 杨光华 Jia Yu;Jin Weiyang;Liu Yang;Chen Yang;Jia Xu;Sun Jie;Tong Yingying;Wang Hua;Yang Guanghua(Shanghai Ocean University:International Research Center for Biological Sciences,Ministry of Science and Technology,Shanghai 201306;Shanghai Ocean University:National Aquatic Animal Pathogen Collection Center,Shanghai 201306;Shanghai Ocean University:Aquatic Animal Genetic Breeding Center,Shanghai 201306;Shanghai Biang Biomedical Technology Co.,Ltd.,Shanghai 201321;Zhejiang Huayi Health Industry Development Co.,Ltd.,Hangzhou 311500,China)

机构地区：[1]上海海洋大学科技部生物科学国际研究中心,上海201306 [2]上海海洋大学国家水生动物病原体收集中心,上海201306 [3]上海海洋大学水产动物遗传育种中心,上海201306 [4]上海比昂生物医药科技有限公司,上海201321 [5]浙江华医健康产业发展有限公司,浙江杭州311500

出　　处：《广东化工》2023年第15期43-46,共4页Guangdong Chemical Industry

基　　金：肿瘤靶向治疗新技术“纳米枪”多中心临床试验(19411951100)。

摘　　要：为改进慢病毒载体生产过程中规模小、生产量不足以及滴度偏低等问题,采用悬浮培养293FT细胞方式,从培养基选择、细胞接种密度、质粒用量等方面对慢病毒包装条件进行优化,通过电击转染并结合生物反应器扩大生产,用离子交换色谱进行纯化以实现慢病毒大规模生产。结果显示以国产培养基M1进行前期细胞扩增培养,用3.75μg/mL质粒电击转染293FT细胞,转染后以2.0×10^(6)/m L的密度接种在优化的2 L生物反应器,用M2培养基培养,最终获得的慢病毒载体滴度达到1.8×10^(12)TU/mL。表明本团队研究的慢病毒载体大规模生产技术可以实现单批次、高滴度、无菌性慢病毒载体的生产,从而为满足临床试验与临床治疗提供基础和保障。To slove the problems of small scale,insufficient production and low titer in the production of lentivirus vectors,we optimized the lentivirus packaging conditions of culture medium selection,cell inoculation density and plasmid dosage,the lentivirus was expanded by electric shock transfection combined with bioreactor,and purified by ion exchange chromatography to achieve large-scale production of lentivirus.The results showed that 293FT cells were transfected with 3.75μg/mL plasmid electroshock and cells were seeded in an optimized 2 L bioreactor at a density of 2.0×10^(6)/mL after transfection,cultured in M2 medium,the lentiviral vector titer reached 1.8×10^(12) TU/mL.In the light of this method could produce a single batch of aseptic lentiviral vector with high titer,thus providing a basis and guarantee for clinical trials and treatment.

关 键 词：慢病毒载体 大规模生产 质量检测 基因治疗 

分 类 号：TQ[化学工程]