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Surface Plasmon Resonance Imaging Measurements of the Inhibition of Shiga-like Toxin by Synthetic Multivalent Inhibitors
journal contribution
posted on 2005-12-01, 00:00 authored by Vishal Kanda, Pavel Kitov, David R. Bundle, Mark T. McDermottA variety of new methodologies to pattern biomolecules
on surfaces and to detect binding events are currently
being developed for high-throughput assay applications.
Carbohydrates serve as attachment sites for toxins, bacteria, and viruses. Immobilized carbohydrate units can
thus be used to directly detect these agents or as a
platform for inhibitor assessment. In this work, modified
glycosides were patterned on gold surfaces to monitor the
binding of the homopentameric B5 cell-recognition subunit of the Shiga-like toxin (SLT). Binding was detected
with the label-free method of surface plasmon resonance
(SPR) imaging. Two synthetic multivalent inhibitors were
used in order to effect inhibitory binding, and SPR
imaging is presented as a simple alternative to ELISA for
the study of toxin inhibition. In contrast to existing
methods for the study of carbohydrate−protein interactions, in particular ELISA, the use of micropatterned
sensor surfaces is shown to be advantageous due to a
decrease in complications and manual labor from numerous blocking, washing, and labeling steps. Carbohydrate
receptor density on the sensor surface was optimized in
order to effect the maximum binding of the SLT. The IC50
values determined were in the low-nanomolar range for
each of the two inhibitors studied.
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Keywords
inhibitor assessmentCarbohydrate receptor densitymethodmicropatterned sensor surfacesImmobilized carbohydrate unitsIC 50 valuesSynthetic Multivalent Inhibitorstoxin inhibitionsensor surfacegold surfacesSurface Plasmon Resonance Imaging MeasurementsELISAbinding eventsSPR imagingsurface plasmon resonancepattern biomoleculesSLTattachment sites
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