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2010全国质谱大会大会报告(一)

2010.8.02

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加拿大Alberta大学 Li, Liang(厉良)教授

  来自加拿大Alberta大学的Li, Liang(厉良)教授,做了题为:“Development of LC/MS Techniques for Comprehensive Metabolome Analysis”(发展LC/MS技术用于综合的代谢组学分析)的报告。在报告中,他简单介绍了代谢组学研究的基本方法、方法的要求,比如代谢组学分析必须要求足够的灵敏度和确定性。而现在面临的挑战有:代谢组学分析中现有技术有限的覆盖率;在定量代谢组学分析中的低通量;鉴定完全未知的代谢物很困难;不知道待研究的某个代谢组到底有多大等等。针对上述挑战,厉良教授课题组发展了一系列LC/MS方法和平台。比如:同位素标记的试剂;有效的LC/MS方法(多维分离技术、三重四极杆、Qtrap等);创建MS/MS数据库;上基于互联网的资源如MycompoundID进行检索;对目标的未知代谢物进行鉴定。以下是英文摘要:

  Metabolomics is a rapidly evolving field for studying biological systems and discovering potential disease biomarkers. For any metabolomics application, metabolome analysis with adequate sensitivity and specificity is essential in defining the metabolome. Ideally, all metabolites present in a biological system are qualitatively and quantitativey profiled. Unfortunately, due to technical limitations, only a fraction of metabolites are currently analyzed by using techniques such as NMR and mass spectrometry (MS). Due to limited metabolome coverage, many important metabolome networks and some subdue changes in the metabolome may not be revealed with current techniques. In this presentation, several technical issues related to the development of LC/MS for enabling metabolome analysis will be discussed.

  Because of great diversity of chemical and physical properties of metabolites, we have been developing an isotope labeling LC/MS workflow with a goal of improving the metabolome coverage in analyzing biological samples such as human biofluids and tissue samples. Several labeling chemistries will be described to provide isotope tags to the metabolites for sensitive detection and accurate quantification. LC method including multi- dimensional separation to separate the labeled metabolites with high efficiency will be discussed. New protocols for MS analysis, metabolite identification and quantitative data processing will be presented.

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赛默飞世尔科技科学仪器市场部经理 王勇为博士

  来自赛默飞世尔科技科学仪器市场部经理王勇为博士主要向大家介绍了Orbitrap质量分析器的工作原理以及该技术的发展现状。

  Orbitrap质量分析器自2000年上市以来,现在已经成为主流的质谱技术。首先,Orbitrap质谱仪特有的关键性能体现在容易操作、并可同时实现超高分辨率(>100000)和高精度质量数。该质量分析器形状如同仿棰体,由仿棰形中心内电极和左右 2 个外仿棰半电极组成。仪器工作时,在中心电极逐渐加上直流高压,在 Orbitrap 内产生特殊几何结构的静电场。当离子进入到 Orbitrap 室内后,受到中心电场的引力,即开始围绕中心电极作圆周轨道运动,同时离子受到垂直方向的离心力和水平方向的推力,而沿中心内电极作水平和垂直方向的震荡。外电极除限制离子的运行轨道范围,同时检测由离子振荡产生的感应电势,其中水平震荡的频率和分子离子的质荷比 (m/z)的平方根成比例关系。

  王博士最后指出,高场Orbitrap和Compact Orbitrap技术的发展使得灵敏度得以提高3倍以上。Orbitrap技术凭借其优异的分析性能使其得到了广泛的应用,应用范围从常规化合物鉴定到复杂基体中痕量化合物分析,如蛋白质组学、药物代谢、药物控制或者食品和饲料中的污染物检测。

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新加坡南洋科技大学 Kai Tang(唐凯)教授

  来自新加坡南洋科技大学的Kai Tang(唐凯)教授,做了题为“Genomic and Proteomic Analyses of Respiratory Viruses”(呼吸病毒症的基因组和蛋白质组分析)的报告。在报告中他谈到:为了更好地了解呼吸道病毒的致病性,课题组用基于质谱分析的技术来表征病毒的基因组序列变异,并发现和病毒作用的宿主蛋白质。比如,他们发展了一套基因组学方法,使被RNaseA酶解的转录产物产生确定位点的断裂,从而有效地应用MALDI分析并鉴定任一单点突变。而他们发展的蛋白质组学方法,可以将病毒以及和宿主细胞作用的蛋白酶解后进行nano-LC-MS/MS分析;被蛋白质组学方法鉴定的宿主蛋白质还可以用不同的生化方法进一步验证。以下是英文摘要:

  In order to better understanding the pathogenicity of respiratory viruses, we have used mass spectrometry based techniques to characterize sequence variations in the virus genome and to discover host proteins that interact with viruses. In the genomics method, virus genome regions between 300 and 800 bases in length were amplified by RT-PCR, followed by in vitro transcription with dU instead of rU or dC instead of rC. The transcription products were then digested with RNase A. Such strategy generated base-specific cleavage products that were ideal for MALDI mass spectrometric analysis. Any single point mutation could be identified by corresponding peak disappearance and appearance of other fragment(s). The method can be particles were purified from their host cells with sucrose gradient. The virus and interacting host cell proteins were solublized in SBS, immobilized and purified in polyacrylamide gel matrix, and then digested in gel. The peptide mixture were extracted and analyzed with nano-LC-MS/MS. Host cell proteins were identified along with known virus proteins after database search and statistical analysis with Trans Proteomic Pipeline. Several host cell proteins indentified with the proteomic approach were further validated with different biochemical assays.


  更多阅读:

  550余人齐聚北国春城 参加质谱大检阅

  赛默飞世尔科技质谱新技术研讨会成功举行

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