Establishment of Single-molecule Enzyme Activity Profiling (SEAP) Platform by Hybrid Technologies of Chemical Biology and Biophysics

Abstract

In this study we have developed a method of profiling multiple “single-molecules” of enzymes in biological samples, by studying their activities as a form of single-molecule enzyme activity assay. The original method for single-molecule enzyme assay in microfabricated chamber devices was reported many years ago, but we for the first time report the application of this concept to identifying each enzyme in the chamber by simultaneously measuring activities against multiple substrates. Based on this idea, we developed the protein profiling technique to globally detect and “count” different sets of enzymes in biological samples containing various characterized and uncharacterized enzymes. We expect that the methodology will open up the application of single-molecule enzyme assay to discovering and using novel biomarker proteins.