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Lectin-Array Blotting

机译：凝集素数阵列印迹

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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.
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