Development of an antibody-lectin enzyme immunoassay for fucosylated α-fetoprotein
Development of an antibody-lectin enzyme immunoassay for fucosylated α-fetoprotein
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DOI：
10.1016/j.bbagen.2011.12.015
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发表时间：
2012-09-01
影响因子：
3
通讯作者：
Taniguchi, Naoyuki
中科院分区：
文献类型：
--
作者：
Korekane, Hiroaki;Hasegawa, Tomoko;Taniguchi, Naoyuki
背景：聚焦化是与癌症相关的最重要的糖基化类型之一。我们之前对肝癌患者甲胎蛋白（AFP）样品的酶基础和结构研究表明，α 1,6-聚焦转移酶（FUT8）的核心聚焦化导致了AFP的聚焦化，事实上，聚焦化的AFP可以用于某些类型的肝硬化肝癌的鉴别诊断。在使用成像技术检测肝癌前3至18个月，这可作为肝癌发展的预测性生物标志物。聚焦AFP目前是通过液相结合测定法（LBA）或电动分析物运输测定法（EATA）来测定的。然而，这些方法需要特殊的仪器，目前只有在主要的医学实验室才能使用。为了克服这一问题，我们尝试建立一种基于特异性抗体和凝集素夹心技术的酶免疫分析法（EIA）。结果：用涂有高浓缩AFP抗体的微滴板检测了人血清中高度浓缩AFP的稀释溶液，该抗体是一种特异性的生物素化金黄念珠菌（AAL）。一种过氧化物酶共轭链亲和素，以及一种化学发光检测系统。该技术能够使用开发的抗体-凝集素EIA结合AFP的富集，在人血清中测量高度浓缩的AFP，稀释至5至80 ng/ml。结论：建立了一种不需要特殊仪器的抗体-凝集素EIA定量浓缩AFP的简便方法。一般意义：该方法可普遍应用于利用特异性抗体定量测定各种集中的糖蛋白。这篇文章是糖蛋白组学特刊的一部分。(C) 2011 Elsevier B.V.版权所有
Background: Fucosylation is one of the most important types of glycosylations related to cancer. Our previous studies of the enzymatic basis and structural studies of alpha-fetoprotein (AFP) samples from liver cancer patients indicated that core-fucosylation by alpha 1,6-fucosyltransferase (FUT8) resulted in the production of fucosylated AFP, and in fact fucosylated AFP allowed differential diagnosis in some types of liver cancer from liver cirrhosis. This served as a predictive biomarker for the development of liver cancer 3 to 18 months before it could be detected using imaging techniques. Fucosylated AFP is currently measured by means of a liquid-phase binding assay (LBA) or by an electrokinetic analyte transport assay (EATA). However, these methods require special instrumentation that is currently available only in major medical laboratories. To overcome this problem, we attempted to develop an enzyme immunoassay (EIA) based on the sandwich technique with specific antibody and lectin.Results: Dilute solutions of highly fucosylated AFP in human sera were assayed using a microtiter plate coated with a periodate-oxidized anti-AFP antibody, a fucose-specific biotinylated Aleuria aurantia lectin (AAL). a peroxidase-conjugated streptoavidin, and a chemiluminescent detection system. The technique was able to measure highly fucosylated AFP diluted to 5 to 80 ng/ml in human sera using the developed antibody-lectin EIA in combination with the enrichment of AFP.Conclusion: A simple method using an antibody-lectin EIA for quantifying fucosylated AFP that does not require special instrumentation was developed.General significance: The method can be generally applied to the quantitative measurement of various fucosylated glycoproteins using specific antibodies. This article is part of a Special Issue entitled Glycoproteomics. (C) 2011 Elsevier B.V. All rights reserved.