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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

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【作者】 秦敏杨国友周富昌涂晓萍廖常如林丽吴先礼

【Author】 QIN Min,YANG Guo-you,ZHOU Fu-chang,et al.(Chengdu Institute of Biological Products, Chengdu 610023,China)

【机构】 成都生物制品研究所成都生物制品研究所 成都610023成都610023成都610023

【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.

【关键词】 甲型副伤寒沙门氏菌LPSO-SP纯化
【Key words】 S.Paratyphoid ALPSO-SPPurification
  • 【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
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