Institute of Bioanalysis, Medical School, University of Pécs, Pécs, Hungary.
János Szentágothai Research Centre, University of Pécs, Pécs, Hungary.
Electrophoresis. 2019 Feb;40(3):447-454. doi: 10.1002/elps.201800378. Epub 2018 Nov 16.
Perchloric acid (PCA) precipitation is a well-known method for the separation of heavily glycosylated proteins and for reducing the masking effect of major serum proteins. The aim of this study is to characterize PCA-soluble serum proteins in healthy individuals and in patients with systemic inflammatory diseases, such as Crohn's disease and sepsis. A PCA precipitation protocol was prepared and adapted to the analytical methods. After PCA treatment of the serum, the soluble proteins in the supernatant were analyzed by SDS-PAGE and by microchip gel electrophoresis (MGE). Characteristic changes of the electrophoretic patterns of the PCA-soluble fractions were observed. Four characteristic bands (at ∼11, ∼65, ∼85, and ∼120 kDa) with varying intensity were detected by MGE. The proportion of the ∼65, ∼85, and ∼120 kDa bands were significantly higher in systemic inflammatory conditions than in healthy individuals (p < 0.001), and characteristic patterns were observed in patients with acute inflammation. The marked differences in the acid-soluble protein patterns, which were observed in patients with ongoing systemic inflammation, might be a good indicator of inflammation. The MGE analysis is a fast screening and quantification method for the detection of characteristic changes among acid-soluble serum proteins.
高氯酸（PCA）沉淀法是一种分离高度糖基化蛋白和减少主要血清蛋白掩蔽效应的常用方法。本研究旨在分析健康个体和系统性炎症性疾病（如克罗恩病和败血症）患者血清中 PCA 可溶蛋白的特性。我们制备了 PCA 沉淀方案并对其进行了调整以适应分析方法。在对血清进行 PCA 处理后，用 SDS-PAGE 和微芯片凝胶电泳（MGE）分析上清液中的可溶性蛋白。观察到 PCA 可溶部分电泳图谱的特征性变化。MGE 检测到四个具有不同强度的特征性条带（约 11、65、85 和 120 kDa）。在系统性炎症条件下，65、85 和 120 kDa 条带的比例显著高于健康个体（p<0.001），并且在急性炎症患者中观察到特征性模式。在持续发生系统性炎症的患者中观察到酸溶性蛋白图谱的显著差异，可能是炎症的良好指标。MGE 分析是一种快速筛选和定量检测酸溶性血清蛋白特征性变化的方法。
