华东理工大学学报（自然科学版）2024，Vol.50Issue(1)：62-70,9.DOI:10.14135/j.cnki.1006-3080.20221112002
新型PCV2衣壳融合蛋白在HEK293F细胞瞬时表达研究
Research on the Transient Expression of a Novel PCV2 Capsid Fusion Protein in HEK293F Cells
罗清平 1彭妍 2ALI Mohsin 1庄英萍 3郭美锦3
作者信息
- 1. 华东理工大学生物反应器工程国家重点实验室,上海 200237
- 2. 上海国佳生化工程技术研究中心有限公司,上海 200237
- 3. 华东理工大学生物反应器工程国家重点实验室,上海 200237||上海国佳生化工程技术研究中心有限公司,上海 200237
- 折叠
摘要
Abstract
Porcine circovirus type 2(PCV2)capsid protein is the only immunodominant structural protein of PCV2 for preparing vaccines against PCV2 infection.The protein can be expressed in Escherichia coli and baculovirus/insect cell expression system.However,studies on the expression of the protein in mammalian cells remain elusive.Here,we investigated several factors that determine the transient expression of the PCV2 capsid protein in mammalian cells.The results showed that a 35%efficiency of the PCV2 capsid gene transfection could be obtained by using HEK293F as the host cell and PEI-40kDa as the transfection reagent.The size and morphology of the PEI/DNA complex was determined by the ratio of the two components,and a particle size range of 15-80 nm contributed to a high cell transfection efficiency.A novel PCV2 Capsid(△1-41aa)-Fc(pig)protein(PCFP)was designed from the immune escape virus strain of PCV2b 41513.PCFP was transiently expressed in HEK293F cells cultivated at a 3 L bioreactor,and the expression level reached 3.8 mg/L.PCFP self-assembled into 41 nm virus-like nanoparticles,which effectively stimulated the humoral immune response in mice.关键词
猪圆环病毒2型/瞬时表达/HEK293F/类病毒颗粒/疫苗Key words
PCV2/transient expression/HEK293F/virus-like particle/vaccine分类
生物科学引用本文复制引用
罗清平,彭妍,ALI Mohsin,庄英萍,郭美锦..新型PCV2衣壳融合蛋白在HEK293F细胞瞬时表达研究[J].华东理工大学学报（自然科学版）,2024,50(1):62-70,9.
