EXPRESSION AND RECOVERY OF RECOMBINANT ROTAVIRUS VP4 FROM INSECT CELL CULTURES

从昆虫细胞培养物中表达和回收重组轮状病毒 VP4

• 批准号: 3803263
• 负责人: M GORZIGLIA
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3803263
• 关键词: DNA virus; Insecta; Rotavirus; gene expression; genetic manipulation; insect virus; microorganism mass culture; protein degradation; protein purification; tissue /cell culture; transfection; virus antigen; virus envelope; virus protein

The possibility of producing and purifying large quantities of recombinant rotavirus VP4 from insect cell cultures was investigated. Spodoptera frugiperda (Sf9) cells were grown in large spinner flasks and infected with a baculovirus expressing the rotavirus OSU VP4 gene. The expressed VP4 subsequently was purified from such cultures to determine the yield of recombinant protein. Conditions for growth of cells and attainment of a high yield of VP4 were optimized to improve the expression and recovery of VP4. The fate of the expressed VP4 antigen and its degradation were examined. Also, a practical purification scheme was developed to maximize the yield of VP4 from insect cell cultures and to reduce cost and complexity of a potential industrial- scale purification process.

生产和提纯大量的 研究了来自昆虫细胞培养物的重组轮状病毒VP 4。 草地贪夜蛾（Spodoptera frugiperda）（Sf 9）细胞在大的转瓶中生长， 用表达轮状病毒OSU VP 4基因的杆状病毒感染。 的 表达的VP 4随后从这样的培养物中纯化以确定 重组蛋白的产量。细胞生长条件和 获得高产量的VP 4进行了优化，以提高 VP 4的表达和恢复。 表达的VP 4抗原的命运 并考察了其降解情况。 此外，一种实用的净化方法 提出了一种从昆虫细胞中最大化VP 4产量的方案 文化，并降低成本和复杂性的潜在工业- 规模纯化工艺。