Microtech2010 2010

Quantum Dot-Peptide Conjugates for Determination of Transferase Enzyme Activity

S.B. Lowe, J.E. Ghadiali, M.M. Stevens
Imperial College London, UK

Keywords: histone acetyltransferase, quantum dot, FRET


We present a quantum dot bio-nanoconjugate system capable of detecting the activity of the histone acetyltransferase, p300, at sub-nM concentrations. CdSe/ZnS core/shell quantum dots were decorated with designer peptides used to mimic the natural substrate of p300; dye-labelled monoclonal antibodies were utilised for detection. In the presence of p300-acetylated peptide, a dye-antibody-peptide-quantum dot complex was formed, facilitating Forster Resonant Energy Transfer (FRET). The quenching of quantum dot emission and corresponding enhancement of dye fluorescence followed, allowing for the quantification of p300 activity. The inhibitor response and limit of detection were also investigated, with sensitivity as low as 600 pM recorded. This quantum dot-based assay presents a rapid, simple alternative to existing enzyme-detection methods. The demonstrated detection sensitivity for p300 is comparable to the current detection standard, radiolabelling. It also provides an excellent response in the biological range and thus has exciting potential applications in medical research and drug discovery.
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