一株无Sf-RV污染的Sf9细胞株生长特性及其在杆状病毒表达系统中的应用  

Growth Characteristics of a Sf9 Cell Line Without Sf-RV Contamination and Its Application in Baculovirus Insect Cell System(BICS)

作　　者：苗丽 侯玉婷 刘发明 朱蕾 吴海华 宋力强 赵雪 杨屹 许雪梅[2] 郭世杰[3] MIAO Li;HOU Yuting;LIU Faming;ZHU Lei;WU Haihua;SONG Liqiang;ZHAO Xue;YANG Yi;XU Xuemei;GUO Shijie(Changchun Zhuoyi Biological Co,.LTD,Changchun 130616,China;Institute of Basic Medical Sciences Chinese Academy of Medical Science,Beijing 100000,China;The First Hospital of Jilin University,Changchun 130021,China)

机构地区：[1]长春卓谊生物股份有限公司,长春130616 [2]中国医学科学院基础医学研究所,北京100000 [3]吉林大学第一医院,长春130021

出　　处：《病毒学报》2020年第6期1151-1156,共6页Chinese Journal of Virology

摘　　要：从雌性草地夜蛾蛹中分离得到的昆虫细胞系(Sf)在杆状病毒昆虫细胞系统(Baculovirus-insect cell system,BICS)中被广泛用作重组蛋白生产的宿主。然而,2014年Ma H等发现该细胞系受到一种病毒的污染,该病毒现在被称为新弹状病毒(Sf-rhabdoviru,Sf-RV)。这提出了一个潜在的安全问题,可能对作为商业重组蛋白生产平台的BICS产生不利影响。因此,我们筛选一种Sf-RV阴性的Sf9细胞株,命名为Sf9-ZY,逆转录-聚合酶链反应(Reverse transcription-polymerase chain reaction,RT-PCR)检测表明,在连续培养的25代过程中,Sf9-ZY细胞没有检测到Sf-RV。Sf9-ZY细胞的一般特性,包括平均生长率、直径,与Sf9细胞基本相同;Sf9-ZY细胞和Sf9细胞产生感染性杆状病毒水平相当;Sf9-ZY和Sf9细胞培养重组杆状病毒表达重组蛋白水平相当。上述结果表明,Sf9-ZY细胞可作为改良的替代宿主,以规避与使用Sf-RV污染的Sf9细胞制造重组蛋白有关的潜在安全危险。Insect cell lines(Sf)isolated from female fall armyworm,Spodoptera frugiperda(Sf),are widely used as hosts for recombinant protein production in baculovirus insect cell system(BICS).However,in 2014,Ma H found that these cell lines were contaminated by a virus now known as Sf-rhabdovirus(Sf-RV).This raises a potential security issue that may adversely affect BICS as a commercial platform for recombinant protein production.Therefore,we screened a Sf-RV-negative Sf9 cell line named Sf9-ZY.Nested RT-PCR detection results showed that Sf-RV was not detected in Sf9-ZY cells during 25 generations of continuous culture.The general characteristics of Sf9-ZY cells,including average growth rate and diameter,are basically the same as Sf9 cells;the level of infectious baculovirus production in Sf9-ZY cells and Sf9 cells is the same level;the level of recombinant baculovirus expression in Sf9-ZY cells and Sf9 cells is the same level.These results suggest that Sf9-ZY cells can be used as an improved alternative host to avoid potential safety risks associated with the production of recombinant proteins by Sf9 cells contaminated with Sf-RV.

关 键 词：SF9细胞 新弹状病毒(Sf-RV) 杆状病毒 细胞单克隆 

分 类 号：Q2-33[生物学—细胞生物学]