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Peptide-nanoparticle hybrid SERS probes for optical detection of protease activity
Authors:Liu Gang L  Rosa-Bauza Yazmin T  Salisbury Cleo M  Craik Charles  Ellman Jonathan A  Chen Fanqing Frank  Lee Luke P
Affiliation:Department of Bioengineering, Biomolecular Nanotechnology Center, Berkeley Sensor and Actuator Center, University of California, Berkeley, CA 94720, USA.
Abstract:Real-time in situ detection of active proteases is crucial for early-stage cancer screening and cell signaling pathway study; however, it is difficult to achieve using fluorescence or radioactive probes at volumes below 1 nL. Here we demonstrated a hybrid optical probe by incorporating nanocrescent particle and peptides with artificial tag molecules. We performed a proof-of-concept study using prostate specific antigen (PSA), one of the most prominent prostate cancer markers, and a serine protease present in patients' seminal fluid and serum. The Raman spectral signal from the tag molecules is enhanced by the nanocrescent and the signal is monitored as the indicator for peptide cleavage in a femtoliter reaction volume, at levels close to a single proteolytically active PSA molecule. The high reaction specificity of the peptides on individual nanoparticles minimizes the false detection of other serine proteases and background Raman signal, which results in a high-fidelity and high-signal-to-noise-ratio cancer nanoprobe that can be easily incorporated into nano/microfluidic devices.
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