不同方法测定基因工程药物中中国仓鼠卵巢细胞蛋白残留量的比较与分析

Comparison and analysis of the CHO cell protein contaminant in bulk of genetic engineering products by using different CHO assays

目的对不同中国仓鼠卵巢 (CHO)细胞蛋白检测试剂盒进行比较分析以确定最佳检测方法。方法用4种不同的CHO细胞蛋白检测试剂对 32批用CHO细胞表达的基因工程产品原液进行检测 ;用标准品替换方法分析 3批红细胞生成素供试品 ;并对 2种CHO细胞蛋白标准品和 4种供试品进行SDS PAGE分析。结果不同的检测方法之间的结果存在较大的偏差 ;同一检测试剂盒替换标准品后测得结果有较大差异 ;标准品的组成成分和比例存在较大差异。

PurposeTo compare and analyze the CHO cell protein contaminant in bulk by using different CHO assays so as to select the ideal method.Methods32 lots of bulk of genetic engineering products which were expressed in CHO cells were detected by different assays. 3 lots of CHO protein in EPO bulk were detected using substitute standards. Two kinds of CHO protein standards and four kinds of samples were analyzed with SDS PAGE.ResultsThe CHO protein content differed from each other sharply. The results were different while we used the same methods but different standards. The components and ratio of two kinds of CHO protein standards also were different.ConclusionDifferent assays should be used for detecting CHO protein content according to the manufacturing and purification process.