48. ELECTROPHORETIC METHOD FOR MONITORING PROTEASE ACTIVITY RESPONSE TO HIGH-FAT MEAL IN PERSONS WITH TYPE 2 DIABETES

Department: Bioengineering
Faculty Advisor(s):

Primary Student
Name: Elaine Alexandra Skowronski
Email: eskowron@ucsd.edu
Phone: 760-717-9237
Grad Year: 2016

Student Collaborators
Augusta Esmeralda Modestino, amodesti@ucsd.edu

Abstract
Proteases have been shown to be elevated in many diseases including shock, diabetes, coagulation disorders, among others. Thus the ability to measure protease activity directly in whole blood will result in better diagnostics, and monitoring of diseases. Current techniques used to measure protease activity required considerable amounts of sample preparation, which is 1) time-consuming 2) costly and 3) alters the sample, making the reads less accurate. We have developed a novel electrophoretic method, which allows rapid measurements of different proteases activities directly in whole blood, requiring no sample preparation, and a single drop of blood. The technology requires charge-changing fluorescent substrates specific to proteases of interests, simple electrophoretic format, and is capable of detecting nanomolar concentrations of protease activity within minutes. Here we present for the first time the detection of MMP-2/-9, elastase and trypsin activities directly in untreated, whole human blood samples of type 2 diabetics (T2D) after a meal. We observed elevated MMP-2/-9 and elastase activities in the blood of T2D compared to normal individuals. We have then demonstrated, that our electrophoretic charge-changing fluorescent substrate assay can be used with real complex clinical samples and no sample preparation for the detection of protease activities to develop better diagnostics and disease monitoring solutions. Moreover, the simplicity of the assay, and the fact that it requires no sample preparation opens the door to the development of point-of-care (POC) system to measure protease activity.

Industry Application Area(s)
Life Sciences/Medical Devices & Instruments

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