HEK293细胞的传代、建库与高效表达重组HPV16-E6E7腺病毒的研究  

Study on HEK293 Cell to Passage, Establish Cell Banks, and Highly Efficiency Express a Recombinant HPV16-E6E7 Adenovirus Vector

作　　者：吴洁[1] 李剑波[1] 高孟[1] 金素凤 潘海桦 陈刚[1] 姜云水[1] 庄昉成[1] 

机构地区：[1]浙江省医学科学院病毒病研究所,杭州310013 [2]浙江普康生物技术股份有限公司,杭州310053

出　　处：《中国现代应用药学》2015年第4期399-403,共5页Chinese Journal of Modern Applied Pharmacy

基　　金：浙江省科技计划项目(2008F3022);杭州市科技局重大科技创新项目(20112313A37)

摘　　要：目的对HEK293细胞进行三级细胞库的建立,并高效表达重组的HPV16-E6E7腺病毒。方法进行HEK293细胞传代培养,并建立三级细胞库,按中国药典2010年版III部要求进行常规检验;培养重组HPV16-E6E7腺病毒,测定滴度,插入目的基因以及基因组酶切图谱鉴定。结果 HEK293细胞传代符合其生长形态,建立的三级细胞库和不同细胞库之间的传代数,细胞浓度和装量均符合疫苗基质制备的要求;能高效表达重组HPV16-E6E7腺病毒,种子滴度>3.0×109 IU·m L?1;表达的目的基因和目的蛋白稳定。结论建立的HEK293三级细胞库符合疫苗用细胞基质的要求。OBJECTIVE To study on HEK293 cell to passage, establish 3-level cell banks, and by that to highly efficiency express a recombinant HPV16-E6E7 adenovirus vector. METHODS HEK293 cell passaged and established 3-level cell banks. And general inspected according to the requirements of Chinese Pharmacopeia Ⅲ 2010. By the HEK293 to propagate a recombinant HPV16-E6E7 adenovirus vector, identitied its titer, inserted target gene and genome macrorestriction. RESULTS HEK293 cell met the growthform of its description; the limited passages, concentration of cells, and install quantity of 3-level cell banks were all conformed to requirements of vaccine substrates; the expressed recombinant HPV16-E6E7 adenovirus titer was more than 3.0×109 IU·m L？1; the expressed inserted target gene and genome macrorestriction map were stable. CONCLUSION HEK293 cell and its 3-level cell banks established are all conformed to the requirements of vaccine substrates.

关 键 词：HEK293 细胞库 HPV16 重组腺病毒 高效表达 

分 类 号：R963[医药卫生—微生物与生化药学]