Title: Biomimetic Surfaces on Microspheres for Protease Diagnostics
Authors: Jose Cornejo1,2, Menake Piyasena2, Jingshu Zhu2, Steven Graves1,2, and Gabriel Lopez1,3
Affiliations: 1Department of Chemical and Nuclear Engineering, University of New Mexico, 2Center for Biomedical Engineering, University of New Mexico, 3Biomedical Engineering Department, Duke University
Abstract: There are many critical diseases that are linked to substances involved in biological pathways such as proteases from viral and bacterial toxins. Investigating such substance – disease interactions are useful for clinical diagnosis, new target drug discovery and delivery, environmental monitoring, and biothreat detection. Some protease –substrate interactions cause severe health risks today. Therefore, it has been of growing interest in recent years to developing inexpensive novel bio-analytical tools that can detect such interactions in high throughput manner. Microsphere based systems enable high sensitive detection due to high surface to volume ratio, thus allowing assembly of high amount of receptors in a small volume. Flow cytometry can be successfully employed in analyzing interactions on biomimetic systems consisting of microspheres bearing supported lipid bilayers. The objective of our current work is to explore novel bio-analytical techniques to investigate protease-substrate interactions on these bioconjugated microspheres using flow cytometry and microfluidic devices. Our overall goal is to develop sensitive, high throughput techniques which will be important in many clinical diagnostic applications. Further software flow cytometric analysis will allow us to quantify and characterize binding, substrate cleavage, and kinetic models. Proteolytic activity leads to substrate cleavage at specific binding sites that will trigger loss of fluorescence intensity of conjugated particles treated with protease. In addition, elucidating such proteolytic mechanisms will contribute to the development of high throughput screening technologies for the discovery of competitive protease inhibitors.