Dual-color proteomic profiling of complex samples with a microarray of 810 cancer-related antibodies.
Dual-color proteomic profiling of complex samples with a microarray of 810 cancer-related antibodies.
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DOI：
10.1074/mcp.m900419-mcp200
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发表时间：
2010-06
期刊：
影响因子：
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通讯作者：
Hoheisel JD
中科院分区：
文献类型：
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作者：
Schröder C;Jacob A;Tonack S;Radon TP;Sill M;Zucknick M;Rüffer S;Costello E;Neoptolemos JP;Crnogorac-Jurcevic T;Bauer A;Fellenberg K;Hoheisel JD
抗体微阵列有潜力对少量样品材料进行全面的蛋白质组分析。这里提出了抗体微阵列在双色模式下的生产、质量评估和可重复应用的方案，具有 1,800 个特征阵列，代表针对 741 种癌症相关蛋白的 810 种抗体。除了阵列质量的测量之外，我们还实施了双色检测的准确性和重要性的指标。在测定重现性和辨别力方面，双色测量优于单色方法。在血清样品的分析中，高丰度蛋白质的消耗并没有提高技术检测质量。相反，耗尽在蛋白质表征中引入了强烈的偏差。在一项初步研究中，我们证明了该方案对尿液样本中蛋白质的适用性。我们发现了胰腺癌患者和健康受试者以及性别之间的尿液样本之间的差异。这项研究表明可以产生生物医学相关数据。正如全面的质量分析所证明的那样，双色抗体阵列方法被证明与其他蛋白质组技术具有竞争力，并且在性能上与转录微阵列分析相当。
Antibody microarrays have the potential to enable comprehensive proteomic analysis of small amounts of sample material. Here, protocols are presented for the production, quality assessment, and reproducible application of antibody microarrays in a two-color mode with an array of 1,800 features, representing 810 antibodies that were directed at 741 cancer-related proteins. In addition to measures of array quality, we implemented indicators for the accuracy and significance of dual-color detection. Dual-color measurements outperform a single-color approach concerning assay reproducibility and discriminative power. In the analysis of serum samples, depletion of high-abundance proteins did not improve technical assay quality. On the contrary, depletion introduced a strong bias in protein representation. In an initial study, we demonstrated the applicability of the protocols to proteins derived from urine samples. We identified differences between urine samples from pancreatic cancer patients and healthy subjects and between sexes. This study demonstrates that biomedically relevant data can be produced. As demonstrated by the thorough quality analysis, the dual-color antibody array approach proved to be competitive with other proteomic techniques and comparable in performance to transcriptional microarray analyses.