新生仔猪大肠埃希氏菌性腹泻K_(88)ac-ST_1-LT_B三价基因工程灭活疫苗生产工艺的研究

Studies on commercial producing process of K_(88)ac-ST_(1)-LT_(B) trivalent gene engineering inactivated vaccine against colibacillus diarrhea of newborn piglets

以实验室工艺为基础,用发酵罐培养,对培养基、诱导剂及诱导条件、通气量、菌液灭活条件等进行了筛选优化。结果表明,重组菌株BL21(DE3)(pXK88acST3LT5)培养的最适培养基为改良LB培养基;乳糖为适合于工业化生产的诱导剂,其最适浓度为100 mmol/L,诱导时间不低于6 h;2×105mL发酵罐培养的最佳通气量为500 L/min;培养菌液的灭活条件为按菌液总量加入4 mL/L甲醛溶液,37℃灭活48 h。按上述工业化生产条件生产疫苗5批,检验结果均安全有效。

On the basis of laboratory process, the culture medium, inducement, ventilation volume and inactivated condition were optimized under industry producing conditions. In consideration of the stability of BL21（DE3） （pXK88acST3LTS） bacterial strain and the cost of production, the improved LB medium was selected as the optimal one. The lactose of 100 mmol/L was the best inducement. In the 200 liter fermenter, the optimal ventilation volume was 500 L/min. The optimal concentration of formaldehyde was 4 mL/L and the inactivating condition was 48 hours at 37 ℃. Five batches of vaccine produced using the optimal commercial conditions were safe and effective, indicating that the commercial producing process was scientifically sound and commercially practical.