脱氧胆酸钠酸沉淀法定量测定b型流感嗜血杆菌结合疫苗中游离多糖的含量  

Quantitative Determination of Free Polysaccharide Content in Haemophilus influenzae Type b Conjugate Vaccine by Acid Precipitation with Sodium Deoxycholate

作　　者：袁军[1] 李新国[1] 瞿明霞[1] 

机构地区：[1]武汉生物制品研究所菌苗室,武汉430207

出　　处：《中国生物制品学杂志》2011年第10期1224-1226,共3页Chinese Journal of Biologicals

摘　　要：目的建立一种b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)结合疫苗中游离多糖含量新的检测方法。方法分别对标准蛋白溶液、多糖溶液和标加衍生多糖(AH-PRP)的结合物原液进行脱氧胆酸钠(NaDC)酸沉淀处理,观察该方法对蛋白和多糖的沉淀效果。分别采用NaDC酸沉淀法和乙醇分步沉淀法测定结合疫苗原液中的游离多糖含量。结果不同浓度的标准蛋白溶液经NaDC酸沉淀法处理后,沉淀中蛋白的回收率在96%～99%之间;不同浓度的多糖溶液经NaDC酸沉淀法处理后,上清中的多糖回收率在99%～106%之间;该方法对结合物中的游离多糖能起到很好的分离效果;两种方法测定结合疫苗原液中的游离多糖含量差异有统计学意义(P<0.01)。结论 NaDC酸沉淀法能专一性地沉淀蛋白物质,对游离多糖无沉淀作用,该方法具有良好的重复性和准确性,可用于测定Hib结合疫苗中的游离多糖含量。Objective To develop a novel method for determination of free polysaccharide content in Haemophilus influenzae type b（Hib） conjugate vaccine.Methods Standard protein solution,polysaccharide solution and bulk of conjugate added with polysaccharide derivative were treated by acid precipitation with sodium deoxycholate（NaDC） and observed for precipitation effect of protein and polysaccharide.The free polysaccharide content in bulk of conjugate vaccine was determined by acid precipitation with NaDC and fractional precipitation with ethanol respectively.Results After acid precipitation with NaDC,the recovery rates of protein in precipitate of standard protein solution at various concentrations were 96% ～ 99%,while those of polysaccharide in supernatant of polysaccharide solution were 99% ～ 106%.The free polysaccharide in conjugate was effectively separated by the developed method.The free polysaccharide contents in bulk of conjugate vaccine determined by acid precipitation with NaDC and fractional precipitation with ethanol showed significant difference（P 0.01）.Conclusion The protein in conjugate was precipitated specifically while no free polysaccharide was precipitated by acid precipitation with NaDC.The developed method showed high reproducibility and accuracy and might used for determination of free polysaccharide content in Hib conjugate vaccine.

关 键 词：嗜血菌 流感 b型 多糖结合疫苗 游离多糖 脱氧胆酸钠 

分 类 号：R378.41[医药卫生—病原生物学] R392-33[医药卫生—基础医学]