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首页> 外文期刊>Proteome science >A new type of protein chip to detect hepatocellular carcinoma-related autoimmune antibodies in the sera of hepatitis C virus-positive patients
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A new type of protein chip to detect hepatocellular carcinoma-related autoimmune antibodies in the sera of hepatitis C virus-positive patients

机译：一种检测丙型肝炎病毒阳性患者血清中与肝细胞癌相关的自身免疫抗体的新型蛋白质芯片

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Background We report here a new type of protein chip to detect antibodies in sera. This chip method was used to a prototype created to detect hepatocellular carcinoma (HCC) -related autoantibodies in the sera of hepatitis C virus (HCV) infected individuals. Results Five cysteine-tagged (Cys-tag) and green fluorescent protein (GFP)-fused recombinant heat shock protein 70 (HSP70), superoxide dismutase 2 (SOD2), and peroxiredoxin 6 (PRDX6), were spotted and immobilized on maleimide-incorporated diamond-like carbon (DLC) substrates. The antibodies in diluted sera were trapped by these proteins at each spot on the chip, and visualized by a fluorescence-conjugated anti-human IgG. The total immobilized protein level of each spot was detected with anti-GFP mouse IgG and a fluorescence-conjugated secondary anti-mouse IgG. The ratio between the two fluorescence intensities was used to quantify autoantibody levels in each serum sample. Heat treatment of the chip in a solution of denaturing and reducing agents, before serum-incubation, improved autoantibody detection. We tested serum samples from healthy individuals and HCC patients using the chips. The HSP70 autoantibodies were found at high levels in sera from HCV-positive HCC patients, but not in HCV-negative sera. Conclusion This protein chip system may have useful properties to capture a specific set of antibodies for predicting the onset of particular cancers such as HCC in HCV-infected individuals.
机译：背景技术我们在此报告一种新型的蛋白质芯片，用于检测血清中的抗体。该芯片方法用于创建原型以检测丙型肝炎病毒（HCV）感染者血清中的肝细胞癌（HCC）相关自身抗体。结果发现并固定了5种半胱氨酸标签（Cys标签）和绿色荧光蛋白（GFP）融合的重组热休克蛋白70（HSP70），超氧化物歧化酶2（SOD2）和过氧化物酶6（PRDX6）。类金刚石碳（DLC）基材。稀释血清中的抗体被这些蛋白质捕获在芯片上的每个位置，并通过荧光偶联的抗人IgG进行可视化。用抗GFP小鼠IgG和荧光缀合的次级抗小鼠IgG检测每个斑点的总固定蛋白水平。两种荧光强度之间的比率用于定量每个血清样品中的自身抗体水平。在血清孵育之前，在变性剂和还原剂溶液中对芯片进行热处理可以改善自身抗体的检测。我们使用该芯片测试了健康个体和HCC患者的血清样本。在HCV阳性HCC患者的血清中发现高水平的HSP70自身抗体，而在HCV阴性血清中则未发现。结论该蛋白芯片系统可能具有捕获特定抗体的有用特性，这些抗体可预测HCV感染个体中特定癌症（例如HCC）的发作。

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