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Ultrasensitive carbohydrate-peptide SPR imaging microarray for diagnosing IgE mediated peanut allergy

机译:超敏糖肽SPR成像微阵列诊断IgE介导的花生过敏

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摘要

Severity of peanut allergies is linked to allergen-specific immunoglobulin E (IgE) antibodies in blood, but diagnostics from assays using glycoprotein allergen mixtures may be inaccurate. Measuring IgEs specific to individual peptide and carbohydrate epitopes of allergenic proteins is promising. We report here the first immunoarray for IgEs utilizing both peptide and carbohydrate epitopes. A surface plasmon resonance imaging (SPRi) microarray was equipped with peptide and β-xylosyl glycoside (BXG) epitopes from major peanut allergen glycoprotein Arachis hypogaea h2 (Ara-h2). A monoclonal anti-IgE antibody was included as positive control. IgEs were precaptured onto magnetic beads loaded with polyclonal anti-IgE antibodies to enhance sensitivity and minimize non-specific binding. As little as 0.1 attomole (0.5 pg/mL) IgE was detected from dilute serum in 45 min. IgEs binding to Ara-h2 peptide and BXG were quantified in 10 μL of patient serum and correlated with standard ImmunoCAP values.
机译:花生过敏的严重程度与血液中的过敏原特异性免疫球蛋白E(IgE)抗体相关,但使用糖蛋白过敏原混合物进行测定的诊断可能不准确。测量对变应原性蛋白质的单个肽和碳水化合物表位具有特异性的IgE很有前景。我们在这里报告利用肽和碳水化合物表位的IgEs的第一个免疫阵列。表面等离振子共振成像(SPRi)微阵列配备了主要花生过敏原糖蛋白花生亚基h2(Ara-h2)的肽和β-木糖基糖苷(BXG)表位。包括单克隆抗IgE抗体作为阳性对照。将IgE预捕获到装有多克隆抗IgE抗体的磁珠上,以增强敏感性并使非特异性结合最小化。在45分钟内从稀血清中检测到低至0.1 attomole(0.5 pg / mL)的IgE。在10μL患者血清中量化与Ara-h2肽和BXG结合的IgE,并将其与标准ImmunoCAP值相关。

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