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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
【摘要】 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。
【Abstract】 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.
- 【文献出处】 微生物学免疫学进展 ,Progress In Microbiology and Immunology , 编辑部邮箱 ,2005年01期
- 【分类号】R392
- 【被引频次】6
- 【下载频次】369