Gold Nanoparticle Based Fluorescence Resonance Energy Transfer Immunoassay for the Detection of the Histone Deacetylase Activity using a Fluorescent Peptide Probe

Abstract

A novel platform for detection of histone deacetylase (HDAC) activity has been developed using a gold nanoparticle based fluorescence resonance energy transfer (FRET) immunoassay. This strategy combined the acetylated fluorescent peptide probe with the anti-acetyl antibody functionalized Au NPs to measure the deacetylation activity of histone deacetylase sirtuin2. Enzymatic deacetylation of the acetylated peptide substrate was detected by a gold nanoparticle labeled anti-acetyl peptide antibody with the formation of the immunocomplex resulting in energy transfer between the fluorescent dyes and the nanoparticles. Due to the highly efficient fluorescence quenching of the gold nanoparticles, the proposed method shows a low background and favorable sensitivity. In addition, this approach can be applied to the evaluation of HDAC inhibitor activity. The proposed platform should facilitate the development of new assays for HDAC activity and other histone modifications.

Keywords:

• Acetylated fluorescent peptide
• Anti-acetyl peptide antibody
• Fluorescence resonance energy transfer (FRET)
• Gold nanoparticles (Au NPs)
• Histone deacetylase (HDAC) activity
• Immunoassay

Acknowledgments

This work was supported by NSFC (21035001, 21275044, 21190041), National Key Basic Research Program (2011CB911000), CSIRT Program, and NSF of Hunan Province (10JJ7002).