冻干ACYW135群脑膜炎球菌多糖结合疫苗中各群多糖含量检测方法的建立及验证

发布时间：2018-06-21 19:43

本文选题：脑膜炎球菌 + 多糖　；参考：《中国生物制品学杂志》2014年08期

【摘要】：目的建立冻干ACYW135群脑膜炎球菌多糖结合疫苗中多糖含量的测定方法。方法将10支ACYW135群脑膜炎球菌多糖结合疫苗用注射用水复溶后,分为10份,70μl/份,依次加入20 mg/ml蛋白酶K溶液0、1、2、3、4、5、6、7、8、9μl,降解结合物中载体蛋白,采用火箭免疫电泳法进行多糖含量的测定,确定蛋白酶K最适加量。对建立的方法进行线性范围、定量限度确定以及准确度、精密性及专属性验证。结果 20 mg/ml蛋白酶K的最适加量为5μl;该方法的线性范围为2.5~40μg/ml,定量限度为2.5μg/ml;3个不同浓度的供试品重复测定3次,A、C、Y、W135群多糖含量的平均回收率分别为99.1%、96.5%、96.5%和97.4%;不同试验人员在不同时间对同一供试品重复测定9次,A、C、Y、W135群多糖含量的CV值分别为3.40%、4.11%、3.35%和4.88%;各群多糖抗原均与其对应的抗血清产生免疫沉淀,而与其他群抗血清不产生免疫沉淀,具有较强的特异性。结论建立了冻干ACYW135群脑膜炎球菌多糖结合疫苗中多糖含量的检测方法,该方法在确定的限度范围内具有较好的准确度、精密性及专属性。
[Abstract]:Objective to establish a method for determining the content of polysaccharide in the lyophilized ACYW135 group meningococcal polysaccharide conjugate vaccine. Methods 10 ACYW135 meningococcal polysaccharide conjugate vaccines were dissolved in 10, 70 mu l/, and 20 mg/ml protease K solution 0,1,2,3,4,5,6,7,8,9 l was added in order to degrade the carrier protein in the conjugate and use rocket The method of immunoelectrophoresis was used to determine the content of polysaccharide and determine the optimum amount of protease K. A linear range, a quantitative limit, accuracy, precision and specificity were used to establish the method. The optimum addition of 20 mg/ml protease K was 5 L; the linear range of the method was 2.5~40 mu g/ml, the quantitative limit was 2.5 u g/ml; 3 different concentrations. The average recovery of the content of A, C, Y and W135 was 99.1%, 96.5%, 96.5% and 97.4%, respectively, for the test samples of A, C, Y and W135, and the CV values of A, C, Y, W135 group polysaccharides were 3.40%, 4.11%, 3.35% and 4.88%, respectively, and the groups of polysaccharide antigens were produced with their corresponding antiserum. Immunoprecipitation and no immunoprecipitation with other group antisera, it has strong specificity. Conclusion the determination method of polysaccharide content in the lyophilized ACYW135 group meningococcal polysaccharide conjugate vaccine is established. The method has good accuracy, precision and specificity in the limits range.
【作者单位】：云南沃森生物技术股份有限公司;
【基金】：云南省科技计划项目(2014AE009)
【分类号】：R392-33;R378.15

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