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Lectin-Array Blotting UNIT 6.12

机译：凝集素阵列印迹单元6.12

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摘要

Aberrant protein glycosylation is a hallmark of cancer, infectious diseases,and autoimmune or neurodegenerative disorders. Unlocking the potential ofglycans as disease markers will require rapid and unbiased glycoproteomicsmethods for glycan biomarker discovery. The present method is a facile andrapid protocol for qualitative analysis of protein glycosylation in complexbiological mixtures. While traditional lectin arrays only provide an averagesignal for the glycans in the mixture, which is usually dominated by the mostabundant proteins, our method provides individual lectin binding profiles forall proteins separated in the gel electrophoresis step. Proteins do not haveto be excised from the gel for subsequent analysis via the lectin array but aretransferred by contact diffusion from the gel to a glass slide presenting multiplecopies of printed lectin arrays. Fluorescently marked glycoproteins are trappedby the printed lectins via specific carbohydrate-lectin interactions and after awashing step their binding profile with up to 20 lectin probes is analyzed witha fluorescent scanner. The method produces the equivalent of 20 lectin blots ina single experiment, giving detailed insight into the binding epitopes presentin the fractionated proteins.
机译：异常蛋白糖基化是癌症，传染病和自身免疫或神经变性障碍的标志。解锁作为疾病标志物的潜在群体将需要快速和无偏糖蛋白质方法进行甘草生物标志物发现。本方法是一种容易性Andrapid方案，可用于复合生物学混合物中蛋白质糖基化的定性分析。虽然传统的凝集素阵列仅为混合物中的聚糖提供的AverignaInm，但是我们的方法在凝胶电泳步骤中提供额外的蛋白质的单个凝集素结合曲线。蛋白质不从凝胶中切除凝胶以通过凝集素阵列从凝胶中切除，但是通过从凝胶的接触扩散到呈现印刷凝集素阵列的多重率的玻璃载玻片中，通过接触扩散来切除。通过特异性碳水化合物 - 凝集素相互作用捕获印刷凝集素的荧光标记的糖蛋白在荧光扫描仪中，荧光扫描仪进行荧光扫描仪后，将印刷的凝胶凝集素捕获。该方法产生相当于20章印迹Ina单一实验，详细介绍结合表位呈现分级蛋白。

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