WARF: P04269US

Protein-Acrylamide Copolymer Hydrogels for Measuring Protein Concentration and Activity


Sean Palecek, Shawn Brueggemeier, Stephen Kron

The Wisconsin Alumni Research Foundation (WARF) is seeking commercial partners interested in developing a method of specifically attaching proteins to glass surfaces.
OVERVIEWProtein microarrays can be used to analyze hundreds or even thousands of proteins in a single experiment. Numerous methods of creating protein microarrays are known; however, no single strategy for attaching proteins to a microarray surface has been widely accepted, in part because current methods suffer from problems such as non-specific protein binding or inaccessibility of attached proteins.
THE INVENTIONUW-Madison researchers have developed a method of specifically attaching proteins to glass surfaces via copolymerization with a polyacrylamide hydrogel. They also have developed techniques for using the arrays formed using this process to detect proteins and measure their concentrations, binding affinities and kinetics.

Their method uses a surface functionalized with an acrylic acid- or acrylamide-based hydrogel. Proteins are labeled with an acrylic moiety and then attached to the functionalized surface through copolymerization with the hydrogel. The attached protein is then available for use in a variety of assays.
  • Creating high density peptide and protein arrays
  • Proteomic studies
  • Biological sensor applications
  • Reduces non-specific surface binding
  • Provides greater access to the attached protein than other methods
  • Proteins are attached in several different orientations, allowing access to different regions.
  • Hydrogel helps maintain the protein in a hydrated, non-denatured state.
  • Porosity of the hydrogel may be varied.
  • Polyacrylamide matrix provides a porous, three-dimensional structure for protein immobilization, greatly increasing the surface area for attachment.
  • Applicable to any protein containing a surface-accessible lysine residue or N-terminus
  • Arrays may be used for kinetic measurements of enzyme reactions, unlike arrays created using other attachment methods.
  • The researchers recently have improved this invention by adding a photocleavable linker at the N-terminus of the protein/peptide, enabling easy, accurate and high throughput detection of compounds in the arrays by matrix-assisted laser desorption ionization (MALDI) mass spectrometry.
STAGE OF DEVELOPMENTUsing this method, the researchers demonstrated that specific attachment of proteins was at least seven-fold greater than non-specific attachment.
Contact Information
For current licensing status, please contact Jennifer Gottwald at or 608-960-9854.
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