通过质谱法进行 DNA 读出的多重蛋白质检测。
Multiplex protein detection with DNA readout via mass spectrometry.
机构信息
Center for Advanced Biotechnology, Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
出版信息
N Biotechnol. 2013 Jan 25;30(2):153-8. doi: 10.1016/j.nbt.2012.11.003. Epub 2012 Nov 28.
Abstract
Multiplex protein quantification has been constrained by issues of assay specificity, sensitivity and throughput. This research presents a novel approach that overcomes these limitations using antibody-oligonucleotide conjugates for immuno-polymerase chain reaction (immuno-PCR) or proximity ligation, coupled with competitive PCR and MALDI-TOF mass spectrometry. Employing these combinations of technologies, we demonstrate multiplex detection and quantification of up to eight proteins, spanning wide dynamic ranges from femtomolar concentrations, using only microliter sample volumes.
摘要
多重蛋白质定量分析一直受到检测特异性、灵敏度和通量等问题的限制。本研究提出了一种新方法，使用抗体-寡核苷酸缀合物进行免疫聚合酶链反应（免疫-PCR）或邻近连接，结合竞争性 PCR 和 MALDI-TOF 质谱，克服了这些限制。通过使用这些技术的组合，我们展示了使用微升级样本量，在从飞摩尔浓度到宽动态范围的范围内，对多达 8 种蛋白质进行多重检测和定量。
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