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应用UPLC与质谱联用技术对5个厂家生产的重组人干扰素理化对照品进行一级结构鉴定及比对研究。通过测定质谱分子量及胰蛋白酶酶解肽图,对其氨基酸序列进行验证,并对二硫键连接方式等翻译后修饰进行分析鉴定。结果 5份对照品的实测分子量均与理论一致;液质肽图结果显示各样品的氨基酸序列均与理论一致;4批样品的二硫键连接方式为Cys1-Cys98、Cys29-Cys138,1批为Cys29-Cys139、Cys86-Cys99;部分样品存在N-末端“+Met”、乙酰化及蛋氨酸氧化等翻译后修饰。液质联用技术可用于干扰素理化对照品的一级结构鉴定及不同厂家产品的比对研究。
Application of UPLC and mass spectrometry technology on the five manufacturers of recombinant human interferon physical and chemical control of the primary structure identification and comparison. The amino acid sequence was verified by measuring the molecular weight of MS and tryptic peptide map, and the post-translational modifications such as disulfide bond were analyzed and identified. Results The measured molecular weights of the five reference products were consistent with the theoretical ones. The results of the liquid chromatogram showed that the amino acid sequences of all the samples were consistent with the theoretical ones. The disulfide bonds of four batches of samples were Cys1-Cys98 and Cys29-Cys138, Cys29-Cys139, Cys86-Cys99; some samples have N-terminal “Met ”, post-translational modification such as acetylation and methionine oxidation. Liquid chromatography-mass spectrometry (LC-MS / MS) can be used to identify the primary structure of interferon reference substance and the comparison between different manufacturers.