Abstract
Protein toxins have been immobilized in a galactoside polyacrylate hydrogel in a microarray format. The large pore size and solution-like environment of these novel hydrogels allow for easy penetration of large proteins and detection reagents. Confocal microscopy provided three-dimensional visualization of dye-labeled toxins cross-linked within the gel and of streptavidin-coated quantum dot (QD) fluorophores used to visualize the toxins after incubation with biotinylated anti-toxin antibodies. Fluorescence microscopy was utilized to visualize arrays of toxins detected by a biotinylated antibody and then exposure to streptavidin-conjugated QDs. The intensity of the QD fluorescence was quantified, and binding to two toxins on three types of hydrogels was examined.
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Acknowledgements
This work was funded by the Office of Naval Research. TJS is supported by a National Research Council Fellowship through the NRL. The views, opinions, and/or findings described in this report are those of the authors and should not be construed as official Department of the Navy positions, policies or decisions.
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Goldman, E.R., O’Shaughnessy, T.J., Soto, C.M. et al. Detection of proteins cross-linked within galactoside polyacrylate-based hydrogels by means of a quantum dot fluororeagent. Anal Bioanal Chem 380, 880–886 (2004). https://doi.org/10.1007/s00216-004-2850-4
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DOI: https://doi.org/10.1007/s00216-004-2850-4