Trypsin Detection Utilizing Peptide Substrates Immobilized on PMMA Nanofibers
We have brought together a unique sensing mechanism and sensing surface for the development of a flow-through sensor for protease detection. Cleavable peptides constructs were designed because of their enhanced performance over antibody systems to detect various analytes. Synthetic
peptides are smaller and are more stable in non-aqueous environments than antibodies and can be designed to be very specific to target analytes. The peptide constructs are immobilized to electrospun PMMA (polymethyl methacrylate) nanofibers, which possess large surface area and allow for 3-dimensional
interaction with the analyte. PMMA is chemically modified leaving a functionalized surface which is then crosslinked to the terminal end of the peptide constructs. The use of a dual-labeled peptide constructs provides the ability to detect protease analytes by monitoring changes in the fluorescence
signal upon cleavage of the constructs by the protease. We have built a sensing system using glass slides and PDMS (polydimethylsiloxane) wells that allow for liquid or gas samples to flow through the sensing system. The sensor system was tested using fluorescence microscopy and a built optical
detection system has been constructed for remote deployment using the nanofiber sensor chips. We have successfully detected trypsin down to 100 pM with response time within 5 minutes. Additional peptide substrates can be designed for detection of many other various analytes and potentially
could help resolve problems with detection equipment used in biothreat detection and medical applications.
Keywords: ELECTROSPUN NANOFIBERS; FLOW-THROUGH SENSING PLATFORM; FLUORESCENCE; PEPTIDE SUBSTRATES; PROTEASE; TRYPSIN
Document Type: Research Article
Publication date: 01 August 2011
- The growing interest and activity in the field of sensor technologies requires a forum for rapid dissemination of important results: Sensor Letters is that forum. Sensor Letters offers scientists, engineers and medical experts timely, peer-reviewed research on sensor science and technology of the highest quality. Sensor Letters publish original rapid communications, full papers and timely state-of-the-art reviews encompassing the fundamental and applied research on sensor science and technology in all fields of science, engineering, and medicine. Highest priority will be given to short communications reporting important new scientific and technological findings.
- Editorial Board
- Information for Authors
- Subscribe to this Title
- Terms & Conditions
- Ingenta Connect is not responsible for the content or availability of external websites
- Access Key
- Free content
- Partial Free content
- New content
- Open access content
- Partial Open access content
- Subscribed content
- Partial Subscribed content
- Free trial content