痢疾结合疫苗LPS制备条件的优化研究

The optimal condition for preparation of LPS of shigella conjugate vaccine

通过对痢疾杆菌LPS提取过程中主要制备环节的优化和改进,确立最佳提取条件和纯化过程,并应用优化后的工艺路线分别制备八批次福氏2 a痢疾杆菌和宋内氏痢疾杆菌LPS;福氏2 a痢疾杆菌LPS批平均产量为1.633g,宋内氏痢疾杆菌LPS批产量平均为1.251g,批产量相对稳定,平均产量比优化改进前提高20%以上。LPS经过酸水解、柱层析纯化获得目标O-SP,福氏2 a痢疾杆菌和宋内氏痢疾杆菌O-SP的得率分别为20%和28%。检测结果证明,LPS和O-SP各项指标均符合规程(草案)相关要求。实验为今后痢疾结合疫苗大规模制备工艺的改进和提高打下了基础。

Optimized the condition of hot phenyl-water LPS extract method.Then prepared 8 lots of Shigella sonnei and Shigella flexneri 2a LPS respectively.The average yield of flexneri 2a LPS is 1.633g and the average yield of sonnei LPS is 1.251g from 200g wet cells.The yield is increased 20%,proteins and nucleic acids content in LPS is less than 5%.After acetic acid hydrolysis and Sephadex G-75 purification,the average yield of flexneri 2a target O-SP antigen is 20% and sonnei target O-SP is 28%,proteins and nucleic acids content in O-SP are both less than 2%.The optimized condition should be solidified in LPS extract protocol and as the base for pilot and large scale procedure.