Biomolecular structural separations by ion mobility-mass spectrometry
Biomolecular structural separations by ion mobility-mass spectrometry
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DOI：
10.1007/s00216-008-1951-x
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发表时间：
2008-06-01
影响因子：
4.3
通讯作者：
McLean, John A.
中科院分区：
文献类型：
--
作者：
Fenn, Larissa S.;McLean, John A.
遵循人类基因组计划的范例，当前的许多系统生物学研究都需要在特定的空间维度（如细胞、组织或有机体水平）和时间维度（如生命周期中的某个点、健康与患病状态）上对样本的生物分子清单进行表征、量化和编目。然后将这些很大程度上与假设无关的数据与生物信息学算法的发展相结合，以获得对不同分子网络如何相互作用和功能的理解，从而为进行假设驱动的研究提供框架[10]。这种简化方法（即测量基因和蛋白质表达水平，评估蛋白质翻译后修饰，代谢物水平等）对于开始参数化调节，控制和扰乱生化过程的异常复杂的分子相互作用网络是必要的，而不是直接分析整个生物实体。基因转录的定量主要是使用基因阵列，而蛋白质表达和代谢物水平通常使用质谱（MS）或色谱分离结合基于MS的技术[2]来测量。通过测量的组合，特定的生物分子网络和疾病状态的分子特征可以被阐明。为了说明此类研究的规模，分别进行了大约105和108次测量，以生成酵母中半乳糖代谢的扰动模型，并确定一套前列腺癌生物标志物[2,3]。这种大规模的实验激发了测量策略的发展，包括更高的吞吐量，更高的选择性，是全面的，并且需要最少的样品操作。基于离子迁移质谱（IM-MS）的二维气相分离技术的新发展为复杂样品的生物分子分离展示了巨大的前景。本趋势报告侧重于IM-MS与纯ms策略相比的几个独特功能，并提供了IM-MS在不久的将来可能发挥越来越大作用的几个关键测量领域的描述。IM-MS技术在生物分子应用方面的详细讨论在其他地方提供[4-9]。
Following the paradigm of the human genome project, much of current systems biology research entails characterizing, quantifying, and cataloging the biomolecular inventory of a sample at specific dimensions of space (eg, cellular, tissue, or organism levels) and time (eg, point in the life cycle, healthy vs. diseased state). These largely hypothesis-independent data are then integrated with the development of bioinformatic algorithms to derive an understanding of how different molecular networks interact and function, which subsequently provide a framework for performing hypothesis-driven studies [1]. This reductionist approach (ie, measuring gene and protein expression levels, evaluating protein post-translational modifications, metabolite levels, etc.) is necessary to begin parameterizing the extraordinarily complex molecular interaction networks that regulate, control, and perturb biochemical processes, rather than directly analyzing the biological entity in its entirety. Quantitation of gene transcription is dominated by the use of gene arrays, while protein expression and metabolite levels are commonly measured using mass spectrometry (MS) or chromatography separations coupled with MS-based techniques [2]. Through a combination of measurements, specific biomolecular networks and molecular signatures of disease-states can be elucidated. To illustrate the scale of such studies, ca. 105 and 108 measurements were performed to generate a perturbation model of galactose metabolism in yeast and to determine a suite of prostate cancer biomarkers, respectively [2, 3]. Such large-scale experiments motivate the development of measurement strategies that incorporate higher throughput, higher selectivity, are comprehensive, and require minimal sample manipulation. Emerging developments in two-dimensional gas-phase separations on the basis of ion mobility–mass spectrometry (IM–MS) have demonstrated great promise for biomolecular separations of complex samples. This trend report focuses on several of the unique capabilities of IM–MS in contrast with MS-only strategies and provides a description of several key measurement areas in which IM–MS will likely play an increasing role in the near future. Detailed discussions of IM–MS techniques for biomolecular applications are provided elsewhere [4–9].