Department of Biochemistry , Purdue University , West Lafayette , Indiana 47907 , United States.
Department of Chemistry , Purdue University , West Lafayette , Indiana 47907 , United States.
Anal Chem. 2018 May 15;90(10):6307-6313. doi: 10.1021/acs.analchem.8b01090. Epub 2018 Apr 23.
Glycoproteins comprise more than half of current FDA-approved protein cancer markers, but the development of new glycoproteins as disease biomarkers has been stagnant. Here we present a pipeline to develop glycoproteins from extracellular vesicles (EVs) through integrating quantitative glycoproteomics with a novel reverse phase glycoprotein array and then apply it to identify novel biomarkers for breast cancer. EV glycoproteomics show promise in circumventing the problems plaguing current serum/plasma glycoproteomics and allowed us to identify hundreds of glycoproteins that have not been identified in blood. We identified 1,453 unique glycopeptides representing 556 glycoproteins in EVs, among which 20 were verified significantly higher in individual breast cancer patients. We further applied a novel glyco-specific reverse phase protein array to quantify a subset of the candidates. Together, this study demonstrates the great potential of this integrated pipeline for biomarker discovery.
糖蛋白占当前美国食品和药物管理局批准的蛋白质癌症标志物的一半以上，但作为疾病生物标志物的新糖蛋白的开发一直停滞不前。在这里，我们通过将定量糖蛋白质组学与新型反相糖蛋白阵列相结合，提出了一种从细胞外囊泡 (EV) 中开发糖蛋白的方案，并将其应用于鉴定乳腺癌的新型生物标志物。EV 糖蛋白质组学在规避当前血清/血浆糖蛋白质组学存在的问题方面显示出巨大的潜力，并使我们能够鉴定出数百种在血液中未被发现的糖蛋白。我们在 EV 中鉴定出 1453 种独特的糖肽，代表 556 种糖蛋白，其中 20 种在个体乳腺癌患者中显著升高。我们进一步应用了新型糖特异性反相蛋白阵列来定量候选物的一部分。总之，这项研究表明，这种综合方案在生物标志物发现方面具有巨大的潜力。
