Detecting acid phosphatase enzymatic activity with phenol as a chemical exchange saturation transfer magnetic resonance imaging contrast agent (PhenolCEST MRI)

Abstract

Phenol contains an exchangeable hydroxyl proton resonant at 4.8 ppm from the resonance frequency of water in the 1H nuclear magnetic resonance (1H NMR) spectrum, enabling itself to be detected at sub-mM concentration by either chemical exchange saturation transfer magnetic resonance imaging (CEST MRI) or exchange-based T2 relaxation enhancement (T2ex) effect under acidic and basic conditions, respectively. We recently investigated the T2ex effects of phenol and its derivatives, but the CEST characteristics of phenols are unknown in detail, and no study on using the natural CEST MRI effects of phenol for detecting enzymatic activity has been conducted. Herein, on the basis of the inherent CEST MR property of phenol, namely phenolCEST, we developed the first MRI approach to detect acid phosphatase (AcP) enzymatic activity. Upon the activity of AcP at pH = 5.0, non-CEST-detectable enzyme substrate phenyl phosphate was converted to CEST-detectable phenol, providing a simple way to quantify AcP activity directly without the need for a second signalling probe. We showed the application of this phenolCEST biosensor for measuring AcP activity in both enzyme solutions and cell lysates of prostate cells. This work opens a door for the utilization of phenolCEST MRI technique in sensor design and development.