Trenchevska Olgica, Schaab Matthew R, Nelson Randall W, Nedelkov Dobrin
The Biodesign Institute at Arizona State University, Tempe, AZ 85287, United States.
The Biodesign Institute at Arizona State University, Tempe, AZ 85287, United States.
Methods. 2015 Jun 15;81:86-92. doi: 10.1016/j.ymeth.2015.02.020. Epub 2015 Mar 7.
The impetus for discovery and evaluation of protein biomarkers has been accelerated by recent development of advanced technologies for rapid and broad proteome analyses. Mass spectrometry (MS)-based protein assays hold great potential for in vitro biomarker studies. Described here is the development of a multiplex mass spectrometric immunoassay (MSIA) for quantification of apolipoprotein C-I (apoC-I), apolipoprotein C-II (apoC-II), apolipoprotein C-III (apoC-III) and their proteoforms. The multiplex MSIA assay was fast (∼ 40 min) and high-throughput (96 samples at a time). The assay was applied to a small cohort of human plasma samples, revealing the existence of multiple proteoforms for each apolipoprotein C. The quantitative aspect of the assay enabled determination of the concentration for each proteoform individually. Low-abundance proteoforms, such as fucosylated apoC-III, were detected in less than 20% of the samples. The distribution of apoC-III proteoforms varied among samples with similar total apoC-III concentrations. The multiplex analysis of the three apolipoproteins C and their proteoforms using quantitative MSIA represents a significant step forward toward better understanding of their physiological roles in health and disease.
先进的快速和广泛蛋白质组分析技术的最新发展加速了蛋白质生物标志物发现和评估的进程。基于质谱（MS）的蛋白质检测方法在体外生物标志物研究中具有巨大潜力。本文描述了一种用于定量载脂蛋白C-I（apoC-I）、载脂蛋白C-II（apoC-II）、载脂蛋白C-III（apoC-III）及其蛋白变体的多重质谱免疫分析（MSIA）方法的开发。该多重MSIA检测方法快速（约40分钟）且高通量（一次可检测96个样本）。该检测方法应用于一小群人类血浆样本，揭示了每种载脂蛋白C都存在多种蛋白变体。该检测方法的定量功能能够单独测定每种蛋白变体的浓度。低丰度蛋白变体，如岩藻糖基化的apoC-III，在不到20%的样本中被检测到。在总apoC-III浓度相似的样本中，apoC-III蛋白变体的分布有所不同。使用定量MSIA对三种载脂蛋白C及其蛋白变体进行多重分析，代表了在更好地理解它们在健康和疾病中的生理作用方面向前迈出的重要一步。
