甲型副伤寒沙门氏菌LPS提取和O-SP纯化

Preparation lipopolysaccharide and purification O-SP from Salmonella paratyphoid A

提取甲型副伤寒沙门氏菌脂多糖(LPS)和纯化特异多糖(O-SP),为制备副伤寒结合疫苗奠定基础.采用大罐培养,菌体热酚法提纯去蛋白,乙醇分级沉淀冷冻离心去核酸,乙酸水解脱毒,高速离心,柱层析等方法纯化.纯化O-SP核酸含量低于1%,蛋白含量低于1.5%,O-乙酰基含量0.5～0.8mmol/L,与甲型副伤寒超免血清形成明显沉淀线,核磁共振图谱(NMR)显示甲型副伤寒O-SP的特征谱.建立了实用的纯化甲型副伤寒O-SP工艺.

To establish a simple method for the purification LPS from S.Paratyphoid A and prepare sufficient amount O-SP for conjugates vaccine research.The strain 50073 was cultured in liquid medium and cells were harvested. We treated the formalin-inactivated cells by hot-phenol to eliminate proteins, alcohol-precipitation to remove nucleic acid and LPS was purified. The LPS was detoxicated by acid hydrolysis, processed by high-speed centrifugation and chromatograph and finally obtained purified O-SP. The purified O-SP contains less than 1% nucleic acid, less than 1.5% protein. O-actyl are 0.5-0.8mmol/g. Double immunodiffusion of LPS and O-SP with hyperimmune serum showed identity reactions. NMR shown the same spectrum as description in the reference.