Three-Dimensionally Functionalized Reverse Phase Glycoprotein Array for Cancer Biomarker Discovery and Validation,Journal of the American Chemical Society

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Three-Dimensionally Functionalized Reverse Phase Glycoprotein Array for Cancer Biomarker Discovery and Validation
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2016-11-17 , DOI: 10.1021/jacs.6b10239
Li Pan , Hillary Andaluz Aguilar , Linna Wang , Anton Iliuk ₁ , W. Andy Tao _{1,

2}

Affiliation

Glycoproteins have vast structural diversity that plays an important role in many biological processes and have great potential as disease biomarkers. Here, we report a novel functionalized reverse phase protein array (RPPA), termed polymer-based reverse phase glycoprotein array (polyGPA), to capture and profile glycoproteomes specifically, and validate glycoproteins. Nitrocellulose membrane functionalized with globular hydroxyaminodendrimers was used to covalently capture preoxidized glycans on glycoproteins from complex protein samples such as biofluids. The captured glycoproteins were subsequently detected using the same validated antibodies as in RPPA. We demonstrated the outstanding specificity, sensitivity, and quantitative capabilities of polyGPA by capturing and detecting purified as well as endogenous α-1-acid glycoprotein (AGP) in human plasma. We further applied quantitative N-glycoproteomics and the strategy to validate a panel of glycoproteins identified as potential biomarkers for bladder cancer by analyzing urine glycoproteins from bladder cancer patients or matched healthy individuals.

中文翻译：

用于癌症生物标志物发现和验证的三维功能化反相糖蛋白阵列

糖蛋白具有广泛的结构多样性，在许多生物过程中起着重要作用，并具有作为疾病生物标志物的巨大潜力。在这里，我们报告了一种新的功能化反相蛋白质阵列 (RPPA)，称为基于聚合物的反相糖蛋白阵列 (polyGPA)，专门用于捕获和分析糖蛋白质组，并验证糖蛋白。用球状羟基氨基树枝状聚合物功能化的硝化纤维素膜用于共价捕获复杂蛋白质样品（如生物流体）中糖蛋白上的预氧化聚糖。随后使用与 RPPA 中相同的经过验证的抗体检测捕获的糖蛋白。我们展示了卓越的特异性、敏感性、通过捕获和检测人血浆中纯化的以及内源性 α-1-酸性糖蛋白 (AGP) 来提高 polyGPA 的定量能力。我们通过分析膀胱癌患者或匹配的健康个体的尿糖蛋白，进一步应用定量 N-糖蛋白组学和策略来验证一组被确定为膀胱癌潜在生物标志物的糖蛋白。

更新日期：2016-11-17

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