Surface immobilized hydrogels as versatile reagent reservoirs for microarrays

J Immunol Methods. 2013 May 31;391(1-2):95-102. doi: 10.1016/j.jim.2013.02.013. Epub 2013 Feb 28.

Abstract

The accuracy of antibody-based microarrays depends on eliminating or at least minimizing the effect of cross-reactive components. In this work, a method to create a versatile, compartmentalized storage system for antibodies and soluble microarray reagents is described. These containers are made of hydrogel plugs immobilized on a slide facing the corresponding sub-array on a printed surface. The hydrogels were polymerized using a 4% w/v N,N-dimethylacrylamide and 3% w/v bisacrylamide solution, thus generating large pore sizes to allow the facile transfer of intact, functional biomolecules. The hydrogel microenvironments can be desiccated and rehydrated with a desired solution, and they can store reagents in a dry form. These hydrogels were shown to provide limits of detection similar to those obtained by conventional incubation conditions in an assay for HIV p24 antigen. Moreover, they prevent cross-reactivity issues in the detection of Staphylococcus aureus enterotoxins A and B and promote the accurate quantification of interleukin 10 by a microarray-integrated calibration curve.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylamides / chemistry*
  • Antibodies / immunology*
  • Antibody Specificity
  • Calibration
  • Cross Reactions
  • Desiccation
  • Enterotoxins / analysis
  • Enterotoxins / immunology
  • Equipment Design
  • HIV Core Protein p24 / analysis
  • HIV Core Protein p24 / immunology
  • Hydrogels*
  • Immunoassay / instrumentation*
  • Immunoassay / standards
  • Interleukin-10 / analysis
  • Interleukin-10 / immunology
  • Limit of Detection
  • Polymerization
  • Porosity
  • Protein Array Analysis / instrumentation*
  • Protein Array Analysis / standards
  • Reproducibility of Results

Substances

  • Acrylamides
  • Antibodies
  • Enterotoxins
  • HIV Core Protein p24
  • Hydrogels
  • Interleukin-10
  • enterotoxin A, Staphylococcal
  • enterotoxin B, staphylococcal
  • N,N-dimethylacrylamide