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PubMed · 16131095

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

Zhenxin Wang·Jason Lee·Andrew R Cossins·Mathias Brust
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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.

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GoldMicroscopy, Atomic ForceNanoparticlesPeptidesProtein Array AnalysisProtein BindingProteinsSolutionsJournal ArticleResearch Support, Non-U.S. Gov't

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Zhenxin Wang, Jason Lee, Andrew R Cossins, Mathias Brust. 2005-09-01. Microarray-based detection of protein binding and functionality by gold nanoparticle probes.. https://doi.org/10.1021/ac050679v

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