Department of Chemical Engineering, Soongsil University, Seoul 06978, Korea.
Analyst. 2019 Mar 25;144(7):2231-2238. doi: 10.1039/c8an02241k.
Serum is one of the most commonly used samples in many studies to identify protein biomarkers to diagnose cancer. Although conventional enzyme-linked immunosorbent assay (ELISA) or liquid chromatography-mass spectrometry (LC-MS)-based methods have been applied as clinical tools for diagnosing cancer, there have been troublesome problems, such as inferior multiplexing capabilities, high development costs and long turnaround times, which are inappropriate for high-throughput analytical platforms. Here, we developed a simple and robust cancer diagnostic method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based total serum protein fingerprinting. First, serum samples were simply diluted with distilled water and subsequently spotted onto a MALDI plate without prior chromatographic purification or separation. The sample preparation method was enough to collect reproducible total serum protein fingerprints and would be highly advantageous for high-throughput assay. Each of the integrated main spectrum profiles (MSPs), which are representative of liver cancer patients (n = 40) or healthy controls (n = 80), was automatically generated by the MALDI Biotyper 3 software. The reliability of the integrated MSPs was successfully evaluated in comparison with a blind test set (n = 31), which consisted of 13 liver cancer patients and 18 healthy controls. Additionally, our partial least squares discriminant analysis (PLS-DA) demonstrated a statistically significant difference in MALDI-TOF MS-based total serum protein fingerprints between liver cancer patients and healthy controls. Taken together, this work suggests that this method may be an effective high-throughput platform technology for various cancer diagnoses and disease evaluations.
血清是许多研究中最常用的样本之一，用于鉴定蛋白质生物标志物以诊断癌症。尽管传统的酶联免疫吸附测定（ELISA）或基于液相色谱-质谱（LC-MS）的方法已被应用于癌症的临床诊断，但仍存在一些问题，例如多重检测能力差、开发成本高、周转时间长等，这些问题使其不适合高通量分析平台。在这里，我们开发了一种简单而强大的癌症诊断方法，该方法基于基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）的总血清蛋白指纹图谱分析。首先，血清样品简单地用蒸馏水稀释，然后直接点样到 MALDI 板上，无需预先进行色谱纯化或分离。这种样品制备方法足以收集可重现的总血清蛋白指纹图谱，并且非常有利于高通量检测。每个集成的主要光谱图谱（MSP），代表肝癌患者（n = 40）或健康对照者（n = 80），均由 MALDI Biotyper 3 软件自动生成。通过与包含 13 名肝癌患者和 18 名健康对照者的盲测试集（n = 31）进行比较，成功评估了集成 MSP 的可靠性。此外，我们的偏最小二乘判别分析（PLS-DA）表明，肝癌患者和健康对照者的 MALDI-TOF MS 总血清蛋白指纹图谱之间存在统计学显著差异。综上所述，这项工作表明，该方法可能是一种用于各种癌症诊断和疾病评估的有效高通量平台技术。
