Microarray-based detection of protein binding and functionality by gold nanoparticle probes

Anal Chem. 2005 Sep 1;77(17):5770-4. doi: 10.1021/ac050679v.

Abstract

We report a microarray format for the detection of proteins and protein functionality (kinase activity) based on marking either specific antibody-protein binding or peptide phosphorylation events by attachment of gold nanoparticles followed by silver deposition for signal enhancement. The attachment of the gold nanoparticles is achieved by standard avidin-biotin chemistry. The detection principle is resonance light scattering. Highly selective recognition of standard proteins (proteins A and G) down to 1 pg/mL for proteins in solution and 10 fg for proteins on the microarray spots is demonstrated. Enzyme activity of the kinase (PKA) is detected with high specificity down to a limit of 1 fg for an established peptide substrate (kemptide) on the microarray spots. Kinase inhibition by the inhibitor (H89) is shown, demonstrating the potential for high-throughput screening for inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gold / chemistry*
  • Microscopy, Atomic Force
  • Nanoparticles / analysis*
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure
  • Peptides / metabolism
  • Protein Array Analysis / methods*
  • Protein Binding
  • Proteins / analysis*
  • Proteins / metabolism*
  • Solutions

Substances

  • Peptides
  • Proteins
  • Solutions
  • Gold