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

Anal Bioanal Chem. 2004 Dec;380(7-8):880-6. doi: 10.1007/s00216-004-2850-4. Epub 2004 Nov 13.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 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

Substances

  • Acrylic Resins
  • Cross-Linking Reagents
  • Galactosides
  • Hydrogels
  • Immunoconjugates
  • Proteins
  • Toxins, Biological
  • carbopol 940