Detection of proteins cross-linked within galactoside polyacrylate-based hydrogels by means of a quantum dot fluororeagent

• Published in: Analytical and bioanalytical chemistry Vol. 380; no. 7-8; pp. 880 - 886
• Main Authors: Goldman, Ellen R, O'Shaughnessy, Thomas J, Soto, Carissa M, Patterson, Jr, Charles H, Taitt, Chris R, Spector, Mark S, Charles, Paul T
• Format: Journal Article
• Language: English
• Published: Germany 01.12.2004
• Subjects: Acrylic Resins - chemistry; Cross-Linking Reagents - chemistry; Fluorescent Antibody Technique; Fluorometry - methods; Galactosides - chemistry; Hydrogels - chemistry; Immunoconjugates - chemistry; Microscopy, Confocal; Microscopy, Fluorescence; Proteins - analysis; Proteins - chemistry; Quantum Dots; Toxins, Biological - analysis; Toxins, Biological - chemistry
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-004-2850-4

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.