Aptamer-based multiplexed proteomic technology for biomarker discovery.
Aptamer-based multiplexed proteomic technology for biomarker discovery.
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DOI：
10.1371/journal.pone.0015004
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发表时间：
2010-12-07
期刊：
影响因子：
3.7
通讯作者：
Zichi D
中科院分区：
文献类型：
--
作者：
Gold L;Ayers D;Bertino J;Bock C;Bock A;Brody EN;Carter J;Dalby AB;Eaton BE;Fitzwater T;Flather D;Forbes A;Foreman T;Fowler C;Gawande B;Goss M;Gunn M;Gupta S;Halladay D;Heil J;Heilig J;Hicke B;Husar G;Janjic N;Jarvis T;Jennings S;Katilius E;Keeney TR;Kim N;Koch TH;Kraemer S;Kroiss L;Le N;Levine D;Lindsey W;Lollo B;Mayfield W;Mehan M;Mehler R;Nelson SK;Nelson M;Nieuwlandt D;Nikrad M;Ochsner U;Ostroff RM;Otis M;Parker T;Pietrasiewicz S;Resnicow DI;Rohloff J;Sanders G;Sattin S;Schneider D;Singer B;Stanton M;Sterkel A;Stewart A;Stratford S;Vaught JD;Vrkljan M;Walker JJ;Watrobka M;Waugh S;Weiss A;Wilcox SK;Wolfson A;Wolk SK;Zhang C;Zichi D
以高度多路复用和高效的方式对蛋白质组（“蛋白质组学”）进行询问仍然是生物学和医学中令人垂涎且具有挑战性的目标。我们提出了一种新的基于适体的蛋白质组学技术，用于生物标志物的发现，能够同时测量来自小样本体积（15微升血清或血浆）的数千种蛋白质。我们目前的检测方法测量了813种蛋白质，检测限较低（中位数为1 pM），总体动态范围为7 log（<100 fM-1 µM），变异系数中位数为5%。该技术通过含有化学修饰的核苷酸的新一代适体实现，其极大地扩展了从中选择适体的大型随机化核酸文库的物理化学多样性。复杂基质（如血浆）中的蛋白质通过将蛋白质浓度特征转化为相应的DNA适体浓度特征的过程进行测量，该过程在DNA微阵列上进行定量。我们的测定利用了适体的双重性质，即具有限定形状的折叠蛋白结合实体和可由特异性杂交探针识别的独特核苷酸序列。为了证明我们的蛋白质组学生物标志物发现技术的实用性，我们将其应用于慢性肾脏疾病（CKD）的临床研究。我们确定了两种众所周知的CKD生物标志物以及另外58种潜在的CKD生物标志物。这些结果证明了我们的技术在快速发现各种疾病状态的独特蛋白质特征方面的潜在效用。我们描述了一个通用的和强大的工具，允许大规模比较离散人群之间的蛋白质组谱。这种无偏见和高度多路复用的搜索引擎将使新的生物标志物的发现方式，是不受我们不完整的生物学知识，从而有助于推进下一代循证医学。
The interrogation of proteomes (“proteomics”) in a highly multiplexed and efficient manner remains a coveted and challenging goal in biology and medicine. We present a new aptamer-based proteomic technology for biomarker discovery capable of simultaneously measuring thousands of proteins from small sample volumes (15 µL of serum or plasma). Our current assay measures 813 proteins with low limits of detection (1 pM median), 7 logs of overall dynamic range (∼100 fM–1 µM), and 5% median coefficient of variation. This technology is enabled by a new generation of aptamers that contain chemically modified nucleotides, which greatly expand the physicochemical diversity of the large randomized nucleic acid libraries from which the aptamers are selected. Proteins in complex matrices such as plasma are measured with a process that transforms a signature of protein concentrations into a corresponding signature of DNA aptamer concentrations, which is quantified on a DNA microarray. Our assay takes advantage of the dual nature of aptamers as both folded protein-binding entities with defined shapes and unique nucleotide sequences recognizable by specific hybridization probes. To demonstrate the utility of our proteomics biomarker discovery technology, we applied it to a clinical study of chronic kidney disease (CKD). We identified two well known CKD biomarkers as well as an additional 58 potential CKD biomarkers. These results demonstrate the potential utility of our technology to rapidly discover unique protein signatures characteristic of various disease states. We describe a versatile and powerful tool that allows large-scale comparison of proteome profiles among discrete populations. This unbiased and highly multiplexed search engine will enable the discovery of novel biomarkers in a manner that is unencumbered by our incomplete knowledge of biology, thereby helping to advance the next generation of evidence-based medicine.
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影响因子：
3.7
作者：
Ostroff RM;Bigbee WL;Franklin W;Gold L;Mehan M;Miller YE;Pass HI;Rom WN;Siegfried JM;Stewart A;Walker JJ;Weissfeld JL;Williams S;Zichi D;Brody EN
通讯作者：
Brody EN
DOI：
10.1111/j.1532-5415.2007.01196.x
发表时间：
2007-06-01
影响因子：
6.3
作者：
Giannelli, Sandra V.;Patel, Kushang V.;Guralnik, Jack M.
通讯作者：
Guralnik, Jack M.
影响因子：
56.9
作者：
Gugliotti, LA;Feldheim, DL;Eaton, BE
通讯作者：
Eaton, BE
影响因子：
158.5
作者：
Gragoudas, ES;Adamis, AP;Guyer, DR
通讯作者：
Guyer, DR
影响因子：
7.8
作者：
Eaton, BE
通讯作者：
Eaton, BE