Development of ultrasensitive lateral flow assays based on horseradish peroxidase enzyme reporters

Lateral flow assays (LFAs), the basis of the home pregnancy test, have been widely used in medical diagnosis, environmental monitoring and forensics. LFAs are attractive point-of care diagnostics due to many reasons, including short test-to-result time, user-friendly format, low cost, storage stability and the ability of non-specialized personnel to run the test. In a typical LFA, gold nanoparticles bearing antibodies to the target are used as the reporter. However, LFA sensitivity lags behind more complex laboratory methods currently used in the clinic. Professor Willson’s group in the Department of Chemical and Biomolecular Engineering (UH) has been developing ultrasensitive LFAs using horseradish peroxidase enzymes (HRP) as the reporter. The primary objective of my summer project was to gain a hands-on experience on how to develop a highly sensitive point-of-care LFA based on HRP enzymes and to evaluate three enzyme substrates using a model LFA.