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Reverse lectin ELISA for detecting fucosylated forms of alpha 1-acid glycoprotein associated with hepatocellular carcinoma

机译:反向凝集素ELISA法检测与肝细胞癌相关的岩藻糖基化形式的α1-酸糖蛋白

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摘要

Altered fucosylation of glycoproteins is associated with development of hepatocellular carcinoma (HCC). Lectins have been commonly used to assay changes in fucosylation of plasma glycoproteins. In the present study a recombinantly engineered form of the fucose binding lectin Aleuria aurantia (AAL) consisting of a single binding site for fucose (S2), was used to construct a reverse lectin ELISA method. Microtiter plates coated with the S2 lectin were used to capture glycoproteins from plasma samples followed by antibody detection of S2-bound fucosylated alpha 1-acid glycoprotein (S2-bound AGP). The method was used to compare the level of S2-bound AGP in serum samples from a small cohort of patients with hepatitis, cirrhosis or HCC. Using the reverse S2 lectin ELISA it was shown that the levels of S2-bound AGP was significantly higher in HCC patients compared to non-cancer patients and that there was also a significant elevation of S2-bound AGP in HCC patients compared to cirrhosis patients. There was no correlation between the level of S2-bound AGP and total AGP concentration. The performance of S2-bound AGP in differentiating HCC from cirrhosis samples or hepatitis samples were compared to other markers. A combination of S2-bound AGP, alpha-fetoprotein and AGP concentration showed performances giving area under receiver operating curves of 0.87 and 0.95 respectively.
机译:糖蛋白岩藻糖基化的改变与肝细胞癌(HCC)的发展有关。凝集素通常被用于测定血浆糖蛋白岩藻糖基化的变化。在本研究中,由岩藻糖(S2)的单个结合位点组成的岩藻糖结合凝集素Aleuria aurantia(AAL)的重组工程形式被用于构建反向凝集素ELISA方法。用涂有S2凝集素的微量滴定板捕获血浆样品中的糖蛋白,然后抗体检测S2结合的岩藻糖基化的α1-酸糖蛋白(S2结合的AGP)。该方法用于比较一小群肝炎,肝硬化或HCC患者血清样本中S2结合型AGP的水平。使用反向S2凝集素ELISA,显示出与非癌患者相比,HCC患者中与S2结合的AGP水平显着更高,并且与肝硬化患者相比,HCC患者中与S2结合的AGP也显着升高。 S2结合的AGP水平和总AGP浓度之间没有相关性。将S2结合的AGP在区分肝癌和肝硬化样品或肝炎样品中的性能与其他指标进行了比较。 S2结合的AGP,甲胎蛋白和AGP浓度的组合显示的性能给出的接收器工作曲线下面积分别为0.87和0.95。

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