Application of an Imprint‐and‐Report Sensor Array for Detection of the Dietary Metabolite Trimethylamine N‐Oxide and Its Precursors in Complex Mixtures

Abstract

Trimethylamine N‐oxide (TMAO) is produced in the gut via metabolism of dietary betaine, choline, and carnitine, and elevated TMAO in plasma is associated with adverse health effects, including cardiovascular events. Currently, we lack high throughput methods for sensing these metabolites and detecting high TMAO. Thus, we have adapted our previously described “imprint‐and‐report” fluorescent sensing method using dynamic combinatorial libraries (DCLs) to create a sensor array for these four metabolites that functions at physiologically relevant concentrations. Templation of DCLs with dye and subsequent addition of analytes generates a fluorescent fingerprint for each metabolite and allows for differentiation via principal component analysis (PCA). Furthermore, we demonstrate that this system can be used to characterize mixtures of the metabolites in both buffer and human plasma samples. Using three to six DCLs, we can distinguish between plasma samples with healthy and elevated levels of TMAO.

Detection and differentiation of per- and polyfluoroalkyl substances (PFAS) in water using a fluorescent imprint-and-report sensor array

“…3). 47,48 The four LCG-templated DCLs formed from these building blocks in 50 mM sodium borate buffer at pH 8.5 (E + G, G + N, E + N, and E + G + N) have been characterized 47 and thus allowed us to gain a facile understanding of the promise of this system for PFAS detection. Despite the building blocks and PFAS all being negatively charged, we anticipated that the fluorinated tails of the PFAS may bind to receptors in aqueous solution due to the hydrophobicity of both species.…”

“… 43 Herein, we describe a novel fluorescent sensor array that differentially detects PFAS, both individual species and complex mixtures in buffer and tap water, at the lowest reported PFAS concentrations (5–40 nM) using dynamic combinatorial chemistry (DCC) 46 to generate a set of dynamic combinatorial libraries (DCLs) containing macrocyclic species templated with a fluorophore. We have previously reported this method, called imprint-and-report sensing, as a high-throughput means for differentiating protein post-translational modifications (PTMs), 47 as well as a set of dietary metabolites in human plasma samples, 48 demonstrating the adaptability and applicability of the system. For the PFAS sensing reported herein, new DCLs containing a fluorinated monomer were optimized to enhance interactions with the fluorinated analytes.…”

“…Moreover, the DCL sensors can readily be adapted for new analytes by modifying the dithiol building blocks that are used. 48 …”