Terracciano Rosa, Gaspari Marco, Testa Flaviano, Pasqua Luigi, Tagliaferri Pierosandro, Cheng Mark Ming-Cheng, Nijdam A Jasper, Petricoin Emanuel F, Liotta Lance A, Cuda Giovanni, Ferrari Mauro, Venuta Salvatore
Department of Experimental and Clinical Medicine, University of Catanzaro Magna Graecia, Catanzaro, Italy.
Proteomics. 2006 Jun;6(11):3243-50. doi: 10.1002/pmic.200500614.
The present manuscript describes a biomarker capturing strategy based on nanoporous silica particles. The method is shown to enrich the yield of species in the low-molecular weight proteome (LMWP), allowing detection of small peptides in the low-nanomolar range. Plasma samples were exposed to the silica particles, and the captured molecular species were profiled using MALDI-TOF. Mass spectra of the silica-treated human plasma samples showed a significant enrichment in MALDI-TOF protein profiles in the LMWP. Preliminary results indicated good level of reproducibility in plasma profiles with CVs on peak heights ranging from 6.3 to 14.7%. The MALDI-TOF signature changed significantly when the characteristics of the nanoporous silica were altered. The facile sample pretreatment before MS analysis, coupled to the potential for tailoring the surface properties of silica supports, hold promise for improving the recovery of low-abundance serum biomarkers.
本手稿描述了一种基于纳米多孔二氧化硅颗粒的生物标志物捕获策略。该方法可提高低分子量蛋白质组(LMWP)中物种的产量,能够检测低纳摩尔范围内的小肽。将血浆样本暴露于二氧化硅颗粒,并用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)对捕获的分子物种进行分析。经二氧化硅处理的人血浆样本的质谱显示,LMWP中的MALDI-TOF蛋白质谱有显著富集。初步结果表明,血浆谱具有良好的重现性,峰高的变异系数(CV)在6.3%至14.7%之间。当纳米多孔二氧化硅的特性改变时,MALDI-TOF特征发生了显著变化。质谱分析前简便的样品预处理,以及定制二氧化硅载体表面性质的潜力,有望提高低丰度血清生物标志物的回收率。