A dynamic range compression and three-dimensional peptide fractionation analysis platform expands proteome coverage and the diagnostic potential of whole saliva.
A dynamic range compression and three-dimensional peptide fractionation analysis platform expands proteome coverage and the diagnostic potential of whole saliva.
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DOI：
10.1021/pr900675w
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发表时间：
2009-12
影响因子：
4.4
通讯作者：
Griffin, Timothy J.
中科院分区：
文献类型：
--
作者：
Bandhakavi, Sricharan;Stone, Matthew D.;Onsongo, Getiria;Van Riper, Susan K.;Griffin, Timothy J.
全面鉴定整个人类唾液中的蛋白质对于了解其全部诊断潜力至关重要。然而，这受到该流体内蛋白质丰度的大动态范围的挑战。为了解决这个问题，我们使用了一个分析平台，该平台将用于动态范围压缩（DRC）的六肽文库与三维（3D）肽分级分离相结合。该方法在整个唾液中鉴定了2340种蛋白质，代表了使用单个分析平台生成的最大唾液蛋白质组数据集。三维肽分馏包括制备IEF，强阳离子交换，和毛细管反相液相色谱的顺序步骤是必不可少的，从DRC的收益最大化。与未用六肽文库处理的唾液相比，DRC显著增加了在物理化学和功能类别中鉴定的蛋白质。大约20%的总唾液蛋白也见于血浆中，两种液体中的蛋白质显示出相当的功能多样性和疾病联系。然而，对于一部分疾病，唾液具有更高的明显诊断潜力。这些结果扩大了全唾液在健康监测/诊断中的潜力，并为改善复杂生物样品的蛋白质组覆盖率提供了一个通用平台。
Comprehensive identification of proteins in whole human saliva is critical for appreciating its full diagnostic potential. However, this is challenged by the large dynamic range of protein abundance within this fluid. To address this problem, we used an analysis platform that coupled hexapeptide libraries for dynamic range compression (DRC) with three-dimensional (3D) peptide fractionation. This approach identified 2340 proteins in whole saliva and represents the largest saliva proteomic dataset generated using a single analysis platform. Three dimensional peptide fractionation involving sequential steps of preparative IEF, strong cation exchange, and capillary reversed phase liquid chromatography was essential for maximizing gains from DRC. Compared to saliva not treated with hexapeptide libraries, DRC substantially increased identified proteins across physicochemical and functional categories. Approximately 20% of total salivary proteins are also seen in plasma, and proteins in both fluids show comparable functional diversity and disease-linkage. However, for a subset of diseases, saliva has higher apparent diagnostic potential. These results expand the potential for whole saliva in health monitoring/diagnostics and provide a general platform for improving proteomic coverage of complex biological samples.
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影响因子：
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作者：
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