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从虚无中挑选消失的蛋白质:如何以可重复且高效的方式收集、运输和共享极稀的蛋白质。

Picking vanished proteins from the void: how to collect and ship/share extremely dilute proteins in a reproducible and highly efficient manner.

作者信息

Bonn Florian, Bartel Jürgen, Büttner Knut, Hecker Michael, Otto Andreas, Becher Dörte

机构信息

Institute for Microbiology, Ernst-Moritz-Arndt University Greifswald , Friedrich-Ludwig-Jahn-Strasse 15, 17487 Greifswald, Germany.

出版信息

Anal Chem. 2014 Aug 5;86(15):7421-7. doi: 10.1021/ac501189j. Epub 2014 Jul 21.

Abstract

Successful proteome analyses of highly dilute samples are strongly dependent on optimized workflows considering especially sample preparation prior to highly sensitive mass spectrometric analysis. Various methods are available for enrichment of proteome samples, each characterized by specific advantages and disadvantages limiting their general application as a method of choice. Here we suggest an optimized universal protocol ensuring reproducibility and effective enrichment of dilute samples by commercial affinity beads. By comparably assessing the performance of the new protocol with selected standard enrichment techniques, we show the seamless application of the enrichment in common mass spectrometry based proteomic workflows. Further, novel applications are suggested including a facile storage and shipping of desiccated, trapped proteome samples at ambient temperatures and usage of the affinity beads for gel-free proteomic approaches.

摘要

对高度稀释样本进行成功的蛋白质组分析在很大程度上依赖于优化的工作流程,尤其要考虑到在高灵敏度质谱分析之前的样本制备。有多种方法可用于蛋白质组样本的富集,每种方法都有其特定的优缺点,这限制了它们作为首选方法的广泛应用。在此,我们提出一种优化的通用方案,通过商业亲和磁珠确保稀释样本的可重复性和有效富集。通过将新方案的性能与选定的标准富集技术进行比较评估,我们展示了该富集方法在基于质谱的常见蛋白质组学工作流程中的无缝应用。此外,还提出了新的应用,包括在环境温度下对干燥、捕获的蛋白质组样本进行便捷的储存和运输,以及将亲和磁珠用于无凝胶蛋白质组学方法。

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