目的 应用串联质谱技术对几种重组蛋白药物的翻译后修饰进行分析。方法 首先将这几种蛋白分别进行蛋白酶切形成多肽片段,然后液相色谱-质谱联用测定其液质肽图,最后应用串联质谱技术分析几种蛋白的翻译后修饰信息。结果 分别对这些蛋白中存在的二硫键、相对分子质量相似肽段、O-糖基化位点以及N-糖基类型等翻译后修饰进行了分析确证。结论 串联质谱技术可在一定程度上弥补蛋白酶切的不足,为重组蛋白药物翻译后修饰的分析提供更加充分的证据。
Abstract
OBJECTIVE To identify several kinds of post-translational modifications in recombinant protein pharmaceuticals by MS/MS. METHODS Firstly the proteins were digested with enzymes, and the obtained peptides were analyzed by UPLC-MS. By MS analysis, peptides with post-translational modifications could be detected, then it was then identified by MS/MS. RESULTS Using MS/MS, the disulfide bonds, peptides with similar molecular mass, O-glycosylation site and the type of N-linked oligosaccharides were identified. CONCLUSION MS/MS analysis can make up for the deficiency of enzyme digestion in some extent, and provide more sufficient evidence for the identification of post-translational modifications.
关键词
串联质谱 /
重组蛋白 /
翻译后修饰 /
糖基化 /
二硫键
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Key words
MS/MS /
recombinant protein /
post-translational modification /
glycosylation /
disulfide bond
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中图分类号：
R917
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脚注
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基金
国家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”资助项目(2012ZX09304010)
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