Systematic discovery of ectopic pregnancy serum biomarkers using 3-D protein profiling coupled with label-free quantitation.
Systematic discovery of ectopic pregnancy serum biomarkers using 3-D protein profiling coupled with label-free quantitation.
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DOI：
10.1021/pr1008866
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发表时间：
2011-03-04
影响因子：
4.4
通讯作者：
Speicher, David W.
中科院分区：
文献类型：
--
作者：
Beer, Lynn A.;Tang, Hsin-Yao;Sriswasdi, Sira;Barnhart, Kurt T.;Speicher, David W.
关键词：
对异位妊娠（EP）和正常宫内妊娠（IUP）的血清蛋白质组进行定量比较，以系统地鉴定候选生物标志物。由丰富的蛋白质免疫耗竭、SDS凝胶、LC-MS/MS和MS信号强度的无标记定量组成的3-D生物标志物发现策略鉴定了70种候选生物标志物，组间差异大于2.5倍。肽量的进一步统计分析用于选择最有希望的12种生物标志物用于进一步研究，其中包括已知的EP生物标志物、新型EP生物标志物（ADAM 12和ISM 2）以及妊娠特异性β-1-糖蛋白家族的5种特异性同种型。技术重复显示出良好的重现性和蛋白质强度，从无标记的发现分析相比，有利的几个已知的参考血清蛋白的丰度水平超过至少三个数量级。类似地，来自无标记发现分析的候选生物标志物的相对丰度与使用用于发现的患者血清池和构成这些池的个体样品的无标记多反应监测对12种最有希望的生物标志物中的5种进行的中试验证测定的相对丰度一致。这些结果表明，强大的，可重复的，深入的3-D血清蛋白质组的发现，随后的中试规模的验证研究，可以很容易地实现使用无标记的定量策略。
Ectopic pregnancy (EP) and normal intrauterine pregnancy (IUP) serum proteomes were quantitatively compared to systematically identify candidate biomarkers. A 3-D biomarker discovery strategy consisting of abundant protein immunodepletion, SDS gels, LC-MS/MS, and label-free quantitation of MS signal intensities identified 70 candidate biomarkers with differences between groups greater than 2.5-fold. Further statistical analyses of peptide quantities were used to select the most promising 12 biomarkers for further study, which included known EP biomarkers, novel EP biomarkers (ADAM12 and ISM2), and five specific isoforms of the pregnancy specific beta-1-glycoprotein family. Technical replicates showed good reproducibility and protein intensities from the label-free discovery analysis compared favorably with reported abundance levels of several known reference serum proteins over at least three orders of magnitude. Similarly, relative abundances of candidate biomarkers from the label-free discovery analysis were consistent with relative abundances from pilot validation assays performed for five of the 12 most promising biomarkers using label-free multiple reaction monitoring of both the patient serum pools used for discovery and the individual samples that constituted these pools. These results demonstrate robust, reproducible, in-depth 3-D serum proteome discovery, and subsequent pilot-scale validation studies can be achieved readily using label-free quantitation strategies.
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