液质联用进行干扰素理化对照品的一级结构鉴定及比对研究.PDFVIP

液质联用进行干扰素理化对照品的一级结构鉴定及比对研究* ** ** 陶磊 裴德宁 韩春梅 陈伟 饶春明 王军志 ( 中国食品药品检定研究院 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) 摘要 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-Cys99、Cys29-Cys139，1 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。 关键词：液质联用 干扰素 理化对照品 一级结构鉴定 比对研究 Characterization and comparison of interferon reference standards using UPLC-MS TAO Lei, PEI De-ning, HAN Chun-mei, CHEN Wei, RAO Chun-ming, WANG Jun-zhi (National Institute for Food and Drug Control, Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products Beijing 100050, China) ABSTRACT: To characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence were verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys99/Cys29-Cys139, 1 lot were Cys29-Cys139/Cys86-Cys99; N-terminal + * 十二五科技重大专项 生物技术药物质量标准和质量控制技术平台（20 12ZX093040 10）”资助项目 **通讯作者 Tel: (010 Fax: (0 10)670519 12; E-mail: raocm@nicpbp.org.cn Tel: (010 Fax: (010)670 14382; E-mail: wangj z@nicpbp.org.cn Met”, acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers. Key words: UPLC-MS interferon refer