Development of a simple and flexible enzyme‐based platform for the colorimetric detection of multiple biomarkers in non‐conventional biofluids

Abstract

Background Enzyme‐based colorimetric systems are inexpensive, simple, adaptable and sensitive methods that allow specific quantification of a substrate. In the clinical field, their use has been the basis for the development of equipment and devices that today are key for disease detection. The objective of this project was to demonstrate the flexibility of a previously developed multi‐enzyme system for colorimetric glucose quantification by adapting and optimizing it for the detection and quantification of other clinically relevant biomolecules, such as galactose, uric acid and 1,5‐anhydroglucitol in buffer conditions. Results The obtained calibration curves for galactose, uric acid and 1,5‐anhydroglucitol show remarkable linearity (R2 ≥ 0.997), precision (CV ≤ 2.38%) and sensitivity with detection and quantification limits of 3.96 and 12.01 μmol L−1; 0.16 and 0.48 μmol L−1; and 0.08 and 0.25 μmol L−1, respectively. In addition, it was found that the three systems are capable of quantifying different concentrations of their respective substrate, showing low variability (CV < 3.3%) and notable recovery percentages (99.21–103.39%). Conclusions The results obtained with the three optimized enzyme‐based colorimetric systems demonstrate the platform flexibility by modifying parameters such as pH buffer, incubation time, enzyme and concentration of the reducing agent under buffer conditions. All results together demonstrate the great potential of this multi‐enzyme platform for the quantification of various substrates in non‐conventional biofluids. © 2022 Society of Chemical Industry (SCI).

3D-Printed Flexible Microfluidic Health Monitor for In Situ Sweat Analysis and Biomarker Detection

“…Due to the low concentration level of the sweat components, the sensing module of wearable sweat sensors is required to demonstrate high sensitivity and excellent selectivity for the quantitative detection of sweat biomarkers. ,, Enzyme-based colorimetric methods have been used in a variety of biosensors over the past decades. − Nanozymes are nanomaterials that mimic natural enzyme properties and possess excellent stability, low cost, easy production, and high catalytic activity, which overcome the drawbacks of natural enzymes. , Among the nanozymes, single-atom catalysts (SACs) with atomically dispersed metal atoms as active sites on their supports have higher catalytic activity and selectivity than traditional nanozymes, endowing the biosensors with higher sensitivity and realizing the sensitive detection of trace biomolecules. , Iron–nitrogen–carbon single-atom catalysts (Fe–N–C SACs) that mimic the natural metalloenzyme structure have well-defined metal atom bond configurations, high level of molecular selectivity, and are easy to fabricate, enabling single-molecule detections with practical-use feasibilities . The Fe–N–C SACs that were synthesized by previous work show excellent peroxidase-like activity. , It has been proven that Fe–N–C SACs can be used in a series of bioassays for ultrasensitive detection of H 2 O 2 , glucose, and amide acid …”