中国动物传染病学报2024，Vol.32Issue(2)：49-57,9.
表达猪瘟病毒E2蛋白的重组杆状病毒反应器工艺研究
Improvement of Bioreactor Process of Recombinant Baculovirus Expressing E2 Protein of Classical Swine Fever Virus
摘要
Abstract
In order to improve the bioreactor process of the recombinant baculovirus expressing classical swine fever virus(CSFV)E2 protein,maximize cell density and increase the expression of CSFV E2 protein,a combined feeding formula was obtained based on the commercial culture medium by optimizing the DOE experimental design of the addition amount of key nutritional components and then the feeding model was optimized on a 3 L bioreactor.The aeration strategy of the bioreactor was further explored to accurately regulate the nutritional and metabolic balances in the culture process of CSFV E2 recombinant baculovirus and solve the problem of the decline of cell viability caused by the excessive accumulation of by-products lactic acid and ammonia.The feeding strategy of semi-continuous feeding process combined with dynamic balance of osmotic pressure and the process mode of directional regulated ventilation were used for the first time to reduce the expression of CSFV E2 protein from 130 mg/L to 520 mg/L,a increase by 3 times.The 3 L bioreactor process was verified in 100 L bioreactor for three consecutive batches.The results showed that the expression of E2 protein was more than 500 mg/L.The vaccine prepared from the harvested CSFV E2 protein induced positive antibody response after 2 weeks of immunization.The antibody level continued to rise and remained at about 80％blocking rate at 8 weeks post inoculation.In summary,the 3 L bioreactor process could be amplified to 100 L bioreactor with the stable expression of CSFV E2 protein and good maintenance of its immunogenicity.关键词
猪瘟病毒/E2蛋白/重组杆状病毒/关键营养单元/半连续补料/定向调节Key words
Classical swine fever virus/E2 protein/recombinant baculovirus/key nutrient units/semicontinuous feeding/ventilation mode分类
农业科技引用本文复制引用
陈晓,李伟国,宋欢欢,苏晓蕊,王同燕,谭菲菲,田克恭..表达猪瘟病毒E2蛋白的重组杆状病毒反应器工艺研究[J].中国动物传染病学报,2024,32(2):49-57,9.基金项目
郑洛新国家自主创新示范区创新引领型产业集群专项(201200211200) （201200211200）
云南省田克恭专家工作站(2019IC062) （2019IC062）
