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用于全局蛋白质组学和唾液生物标志物发现的自组装 STrap。

Self-Assembled STrap for Global Proteomics and Salivary Biomarker Discovery.

机构信息

J. Craig Venter Institute , 9605 Medical Center Drive , Rockville , Maryland 20850 , United States.

J. Craig Venter Institute , 4120 Capricorn Lane , La Jolla , California 92037 , United States.

出版信息

J Proteome Res. 2019 Apr 5;18(4):1907-1915. doi: 10.1021/acs.jproteome.9b00037. Epub 2019 Mar 21.

Abstract

Clinical biomarkers identified by shotgun proteomics require proteins in body fluids or tissues to be enzymatically digested before being separated and sequenced by liquid chromatography-tandem mass spectrometry. How well peptide signals can be resolved and detected is largely dependent on the quality of sample preparation. Conventional approaches such as in-gel, in-solution, and filter-based digestion, despite their extensive implementation by the community, become less appealing due to their unsatisfying protein/peptide recovery rate, lengthy sample processing, and/or lowcost-effectiveness. Suspension trapping has recently been demonstrated as an ultrafast approach for proteomic analysis. Here, for the first time, we extend its application to human salivary proteome analyses. In particular, we present a simple self-assembled glass fiber filter device which can be packed with minimal difficulty, is extremely cost-effective, and maintains the same performance as commercial filters. As a proof-of-principle, we analyzed the whole saliva from 8 healthy individuals as well as a cohort of 10 subjects of oral squamous cell carcinoma (OSCC) patients and non-OSCC subjects. Label-free quantification revealed surprisingly low interindividual variability and several known markers. Our study provides the first evidence of an easy-to-use and low-cost device for clinical proteomics as well as for general proteomic sample preparation.

摘要

通过鸟枪法蛋白质组学鉴定的临床生物标志物需要将体液或组织中的蛋白质在通过液相色谱-串联质谱进行分离和测序之前进行酶消化。肽信号的分辨率和检测效果在很大程度上取决于样品制备的质量。尽管传统方法(如胶内、溶液内和基于滤器的消化)已被广泛应用于社区,但由于其蛋白质/肽回收率低、样品处理时间长和/或成本效益低,其吸引力降低。悬浮捕获最近已被证明是一种用于蛋白质组分析的超快方法。在这里,我们首次将其应用扩展到人类唾液蛋白质组分析中。具体来说,我们提出了一种简单的自组装玻璃纤维滤器装置,该装置易于填充,成本效益极高,并且与商业滤器具有相同的性能。作为原理验证,我们分析了 8 名健康个体以及 10 名口腔鳞状细胞癌(OSCC)患者和非 OSCC 患者的全唾液。无标记定量分析显示出令人惊讶的低个体间变异性和几个已知的标志物。我们的研究首次提供了一种易于使用且成本低廉的临床蛋白质组学设备以及一般蛋白质组样品制备的证据。

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