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目的 探讨表达人源抗HBsAgFab工程化大肠杆菌的优化培养模式及诱导方法 ,以期大量获取人源抗HBsAgFab。方法 在摇瓶发酵条件下探讨工程菌的生长和表达规律 ,筛选最佳的培养模式及诱导表达条件 ,后于发酵罐中行补料高密度发酵试验 ,以确定最佳补料模式。结果 由摇瓶发酵获得的数据表明 ,当培养体系处于对数生长中期为诱导表达的起点 ,在 2 5℃下以 0 2 %arabinose诱导12h条件下Fab的产率最佳 ,采用溶氧控制补料模式可使培养体系的最大OD60 0 达到 5 5 2 ,相当于湿菌含量 110g/L水平。同时 ,经证实Fab的抗原结合活性良好。结论 确定了周期短、产率高且稳定可靠的发酵工艺 ,为应用原核表达体系工业化大批量生产基因工程抗体奠定了基础
Objective To investigate the optimal culture mode and induction method of humanized anti-HBsAgFab engineered Escherichia coli in order to obtain large quantities of human anti-HBsAgFab. Methods The growth and expression of engineered bacteria were studied under shake flask fermentation conditions. The best culture mode and induced expression conditions were screened, and then fed-batch high-density fermentation was conducted in the fermentor to determine the optimal feeding mode. Results The data obtained from shake flask fermentation showed that when the culture system was at the midpoint of logarithmic growth phase, the yield of Fab was best under the induction of 0 2% arabinose at 25 ° C for 12 hours, Material model allows the maximum OD60 0 of the culture system to reach 5 5 2, which corresponds to a level of 110 g / L of wet bacteria. Meanwhile, the antigen-binding activity of Fab was confirmed to be good. Conclusion The conclusion is drawn that the fermentation process with short cycle, high yield, stable and reliable, laid the foundation for the industrialized mass production of genetically engineered antibody in prokaryotic expression system