Surface Plasmon Resonance Imaging Measurements of the Inhibition of Shiga-like Toxin by Synthetic Multivalent Inhibitors

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• 作者：Vishal Kanda ; Pavel Kitov ; David R. Bundle ; and Mark T. McDermott
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：December 1, 2005
• 年：2005
• 卷：77
• 期：23
• 页码：7497 - 7504
• 全文大小：306K
• 年卷期：v.77,no.23(December 1, 2005)
• ISSN：1520-6882

文摘

A variety of new methodologies to pattern biomoleculeson surfaces and to detect binding events are currentlybeing developed for high-throughput assay applications.Carbohydrates serve as attachment sites for toxins, bacteria, and viruses. Immobilized carbohydrate units canthus be used to directly detect these agents or as aplatform for inhibitor assessment. In this work, modifiedglycosides were patterned on gold surfaces to monitor thebinding of the homopentameric B₅ cell-recognition subunit of the Shiga-like toxin (SLT). Binding was detectedwith the label-free method of surface plasmon resonance(SPR) imaging. Two synthetic multivalent inhibitors wereused in order to effect inhibitory binding, and SPRimaging is presented as a simple alternative to ELISA forthe study of toxin inhibition. In contrast to existingmethods for the study of carbohydrate-protein interactions, in particular ELISA, the use of micropatternedsensor surfaces is shown to be advantageous due to adecrease in complications and manual labor from numerous blocking, washing, and labeling steps. Carbohydratereceptor density on the sensor surface was optimized inorder to effect the maximum binding of the SLT. The IC₅₀values determined were in the low-nanomolar range foreach of the two inhibitors studied.