口蹄疫灭活疫苗中蛋白质含量测定方法的比较

Comparison of Protein Quantitation Assaysinquality test of FMD inactivated vaccine

关键词:  口蹄疫灭活疫苗  蛋白质含量测定  凯氏定氮法  改良Lowry法  考马斯亮蓝法  SDS-PAGE

Key words:  FMD inactivated vaccine  protein quantitation  Kjeldahlnitrogen method  Modified Lowryprotein assay  Coomassie Plus assay  SDS-PAGE

摘要:

口蹄疫灭活疫苗中蛋白质含量的测定,可以反应出疫苗生产工艺的优劣,可间接反应疫苗质量。采用凯氏定氮法,改良Lowry法和考马斯亮蓝法三种较常见的蛋白质含量测定方法,对全国六个不同口蹄疫灭活疫苗生厂家共计15批疫苗的总蛋白质含量进行测定,并对测定结果进行比较,同时采用SDS-PAGE对样品中蛋白质含量进行定性，验证以上三种方法测定结果。试验结果表明,这些疫苗的总蛋白质含量差异比较大,1ml水相样中蛋白质含量在50～2000μg范围内。三种检测方法比较表明,改良Lowry法能较真实的反映口蹄疫灭活疫苗中蛋白质含量,与凯氏定氮法相比,此方法操作简单,工作效率高,而且低耗。

Abstract:

The Protein quantitationofFMD inactivated vaccine,couldevaluate the vaccine production process and indirectly reflect the purity of vaccine. In this study,three protein quantitation assays, Kjeldahlnitrogen method,Modified Lowryprotein assay and Coomassie Plusassay, were used to quantitate protein of 15 batches of FMD inactivated vaccine from six FMD inactivated vaccinemanufacturersin China respectively,then thedata were compared，and the Protein qualitative analysis of SDS-PAGE were used to validate the data. These results indicated that there was significant difference in protein content of these vaccines,and theprotein content ranged from 50ug to 2000ug in 1ml aqueous sample.In compared with these three methods,Modified Lowryprotein assay could perform protein quantitationmore truly,and compared to the Kjeldahl method, this method is simple, efficient, and low consumption.