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2. 发明授权

US06391649B1 Method for the comparative quantitative analysis of proteins and other biological material by isotopic labeling and mass spectroscopy 有权
标题翻译：通过同位素标记和质谱法对蛋白质和其他生物材料进行比较定量分析的方法
公开(公告)号：US06391649B1
公开(公告)日：2002-05-21
申请号：US09304799
申请日：1999-05-04
申请人： Brian T. Chait , David Cowburn , Yoshi Oda
发明人： Brian T. Chait , David Cowburn , Yoshi Oda
IPC分类号： G01N3100
CPC分类号： G01N33/6842 , G01N33/6803 , Y10T436/24 , Y10T436/25 , Y10T436/25375
摘要： The present invention is a method for accurately comparing the levels of cellular components, such as proteins, present in samples which differ in some respect from each other using mass spectroscopy and isotopic labeling. A first sample of biological matter, such as cells, is cultured in a first medium and a second sample of the same biological matter is cultured in a second medium, wherein at least one isotope in the second medium has a different abundance than the abundance of the same isotope in the first medium. One of the samples is modulated, such as by treatment with a bacteria, a virus, a drug, hormone, a chemical or an environmental stimulus. The samples are combined and at least one protein is removed. The removed protein is subjected to mass spectroscopy to develop a mass spectrum. A ratio is computed between the peak intensities of at least one closely spaced pair of peaks to determine the relative abundance of the protein in each sample. The protein is identified by the mass spectrum or through other techniques known in the art. Modifications to the proteins, such as the phosphorylation of the protein, and the site of the modification may also be determined through the process of the present invention. The method is applicable to the components of any type of biological matter which are ionizable and may therefore be analyzed by mass spectroscopy.
摘要翻译：本发明是用于使用质谱和同位素标记来精确比较在某些方面彼此不同的样品中存在的样品中蛋白质等细胞成分水平的方法。生物物质如细胞的第一样本在第一培养基中培养，同一生物物质的第二个样品在第二培养基中培养，其中第二培养基中的至少一个同位素具有与丰度不同的丰度第一种培养基中的相同同位素。其中一个样品被调节，例如通过用细菌，病毒，药物，激素，化学或环境刺激物治疗。将样品合并并除去至少一种蛋白质。将去除的蛋白质进行质谱以产生质谱。在至少一个紧密间隔的峰对的峰值强度之间计算比率，以确定每个样品中蛋白质的相对丰度。蛋白质通过质谱或本领域已知的其它技术鉴定。蛋白质的修饰，如蛋白质的磷酸化和修饰的位点也可以通过本发明的方法来确定。该方法适用于可离子化的任何类型生物物质的组分，因此可通过质谱分析。

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