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

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 (R-2 >= 0.997), precision (CV <= 2.38%) and sensitivity with detection and quantification limits of 3.96 and 12.01 mu mol L-1; 0.16 and 0.48 mu mol L-1; and 0.08 and 0.25 mu mol 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. (c) 2022 Society of Chemical Industry (SCI).

Automated summary

The study demonstrates the flexibility of an enzyme-based colorimetric system for the accurate quantification of various clinically relevant biomolecules. The optimized system shows excellent linearity, precision, and sensitivity, with low variability and high recovery rates. This multi-enzyme platform has great potential for quantifying different substrates in non-conventional biofluids.