同位素亲和标签_基质辅助激光解吸电离飞行时间串联质谱

基质辅助激光解吸电离飞行时间质谱用于蛋白质相对定量方法的研究

http://www.100md.com 《分析化学》 2006年第7期

同位素亲和标签,基质辅助激光解吸电离飞行时间串联质谱,毛细管反相液相色谱,,同位素亲和标签,基质辅助激光解吸电离飞行时间串联质谱,毛细管反相液相色谱,基质辅助激光解吸电离飞行时间质谱用于蛋白质相

     摘要 将亲和标记技术和基质辅助激光解吸电离飞行时间串联质谱联用，建立了蛋白质相对定量方法，以牛血清白蛋白为实验对象，考察了该方法的准确度、重现性等指标;又以数种标准蛋白混合物为研究对象，考察了该方法的动态范围及相应的标准偏差，为实际生物样品中差异蛋白质分析奠定了基础。

    关键词 同位素亲和标签,基质辅助激光解吸电离飞行时间串联质谱,毛细管反相液相色谱

    Studies on Relative Quantitative Method for Differential Oroteomics byIsotopecoded Affinity Tag and Matrixassisted Laser Desorption IonizationTime of FlightTime of Flight Mass Spectrometry

    Meng Qingfang, Zhang Yangjun, Cai Yun1, Qian Xiaohong

    (Department of Genomics and Proteomics, Beijing Institute of Radiation Medicine, Beijing 100850)

    (Beijing Proteome Research Center, Beijing 102206)

    Abstract A relative quantitative method for differential proteomics by cleaving isotope coded affinity tag (cICAT) and matrixassisted laser desorptin ionization ionizattion time of flihgt fandem mass spectrometry (MALDTOFTOFMS) was established. The accuracy and reproducibility of the method were evaluated by bovine serum albumin digest with relative standard deviation less than 30% and good reproducibility. Dynamic range was also evaluated by analyzing two mixtures of several standard proteins with different dynamic range respectively. The experimental results showed that in the dynamic range of 1∶30, the quantitation error was less than 30%. Although the quantitation error becomes very large when beyond this range, it does not affect getting the information of differential proteins. All the works provide an alternative method for differential proteomics analysis in biological samples. ......

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