两种Lowry法检测百日咳疫苗原液蛋白含量的比较  

Comparison of two Lowry methods for determination of protein content in bulk of Boredella pertussis

作　　者：卫辰[1,2] 骆鹏[2] 王丽婵[2] 张华捷[2] 晁哲[2] 吴燕[2] 侯启明[2] 马霄[2] 胡云章[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物研究所,云南昆明650118 [2]中国食品药品检定研究院百白破疫苗与毒素室卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中国生物制品学杂志》2017年第5期519-524,共6页Chinese Journal of Biologicals

基　　金：国家863计划"疫苗效果和质量评价新技术研究"(2012AA02A402)

摘　　要：目的比较《中国药典》三部(2015版)收录的两种Lowry法测定百日咳疫苗原液蛋白定量的差异,为原液蛋白准确定量和新版药典Lowry法的适用性确认提供依据。方法 9个实验室分别按照现行药典收录的两种Lowry法,对4家15批百日咳疫苗原液蛋白含量进行测定。并与凯氏定氮法检测结果进行比较。结果方法 1(未经酸沉淀Lowry法)实验室内测定结果的变异系数(CV)均值在1.7%~8.9%之间,实验室间测定结果的CV值在3.9%~9.3%之间;方法 2(酸沉淀Lowry法)实验室内测定结果的CV均值在3.6%~7.4%之间,实验室间测定结果的CV值在3.9%~18.1%之间。同一样品两种方法测定结果差异有统计学意义(P<0.001),结果呈相关性(R2=0.944 9)。在与部分样品测得的凯氏定氮法结果的比较中,方法 1差异有统计学意义(P<0.001),方法 2差异无统计学意义(P>0.05)。结论所有样品采用同一种Lowry方法测定时所得结果实验室内和实验室间一致性较好,采用方法 2测定结果明显低于方法 1,更接近凯氏定氮法结果真实值,表明酸沉淀对Lowry法百日咳原液蛋白定量有显著影响。与Lowry方法 1相比,Lowry方法 2更能准确反映百日咳疫苗原液的真实蛋白含量,且精密度、适用性均符合要求。Objective To investigate difference between two Lowry methods in Chinese Pharmacopeia （Volume Ⅲ,2015 edition） for determination of protein content in bulk of Boredella pertnssis,and provide a basis for accurate quantitation of protein content in the bulk and confirmation of suitability of Lowry method in the new pharmacopoeia.Methods Fifteen batches of bulks of B.pertussis from four manufacturers were determined for protein content by nine laboratories with two Lowry methods in Chinese Pharmacopeia （Volume Ⅲ,2015 edition）,of which the results were compared with those by Kjeldahl nitrogen determination method.Results The mean CVs of results of intra-assays by method 1 （Lowry method without acid precipitation） by various laboratories were 1.7% ～ 8.9%,while those of inter-assays were 3.9% and 9.3% respectively.However,the mean CVs of results of intra-and inter-assays by method 2 （Lowry method with acid precipitation） were 3.6% ～ 7.4% and 3.9% ～ 18.1% respectively.The determination results of the identical samples by the two methods showed significant difference （P 〈 0.001） and a correlation （R2 =0.944 9）.The determination results of partial samples by method 1 showed significant difference with that by Kjeldahl method （P 〈 0.001）,while those by method 2 showed no significant difference （P 〉 0.05）.Conclusion All the results of samples determined the same Lowry method showed good consistence in both intra-and inter-assays.However,the determination results by method 2 were significantly lower than those by method 1,and were closer to the true values of determination results by Kjeldahl method,indicating significant effect of acid precipitation on quantitative determination results of protein content in bulk of B.pertussis by Lowry method.Compared with method 1,method 2 reflected the protein content in bulk of B.pertussis more accurately,of which the precision and suitability met the relevant requirements.

关 键 词：百日咳 原液 蛋白定量 Lowry法 

分 类 号：R392-33[医药卫生—免疫学]