Detection of proteins cross-linked within galactoside polyacrylate-based hydrogels by means of a quantum dot fluororeagent.
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.
Goldman
Ellen R
ER
U.S. Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, Washington, DC 20375, USA. egoldman@cbmse.nrl.navy.mil
O'Shaughnessy
Thomas J
TJ
Soto
Carissa M
CM
Patterson
Charles H
CH
Taitt
Chris R
CR
Spector
Mark S
MS
Charles
Paul T
PT
D000180:Acrylic Resins / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D005470:Fluorometry / Q000379:methods; D005697:Galactosides / Q000737:chemistry*; D020100:Hydrogels / Q000737:chemistry*; D018796:Immunoconjugates / Q000737:chemistry; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D045663:Quantum Dots; D014118:Toxins, Biological / Q000032:analysis / Q000737:chemistry
D000180:Acrylic Resins; D003432:Cross-Linking Reagents; D005697:Galactosides; D020100:Hydrogels; D018796:Immunoconjugates; D011506:Proteins; D014118:Toxins, Biological; C006903:carbopol 940