Mr. Craig R. Weilbaecher Profile

Craig R. Weilbaecher

at Univ of Missouri/Columbia

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Proceedings Article | 4 October 2007 Paper

Development of a novel nanomaterial-based optical platform for a protease biosensor

Craig Weilbaecher, Maruf Hossain, Shubhra Gangopadhyay, Sheila Grant

Proceedings Volume 6759, 67590D (2007) https://doi.org/10.1117/12.734679

KEYWORDS: Nanoparticles, Gold, Luminescence, Thin films, Biosensors, Molecules, Thin film deposition, Biomedical optics, Physical vapor deposition, Fluorescence resonance energy transfer

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We have been investigating a new nanomaterial-based optical platform for the immobilization of protease substrates for the development of a biosensor to detect medically relevant enzymes. Metallic nanoparticles have been deposited onto thin films and are being used for their optical properties. Two different peptide sequences have been designed as trypsin substrates that are designed to be immobilized onto the surface of the thin films. The peptides were synthesized with a fluorophore attached at the terminal end of the peptide to allow for fluorescence sensing. Fluorescent molecules in close proximity to metallic elements will have their fluorescence signal quenched due to surface plasmon resonance (SPR) effects. When the peptide is cleaved by trypsin, the fluorophore is separated, resulting in a detectable change in fluorescence intensity. These novel nanomaterial-based optical platforms have been fabricated using physical vapor deposition. Innovative techniques have been invented using these machines to acquire nanoparticles in the range of a few nanometers on these thin films. It is known that nanoparticles with dimensions less than their bandwidth display optical properties much different from their bulk counterparts. We have immobilized the peptide substrates to the surface of the metallized thin films so they are in close proximity with each other. Polydimethlysiloxane (PDMS) was molded to create small wells and placed on the thin films. Fluorescent microscopy was used to image the wells as various concentrations of the enzyme were introduced resulting in a recovery of green fluorescence from the fluorophore on the cleaved portion of the peptide. Different size nanoparticles and different immobilization processes are being used to optimize the design of the protease biosensor.

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