| Abstract
| - 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.
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