Abstract
A new nano-structured platform for fluorescent analysis using PQQ-dependent glucose dehydrogenase (PQQ-GDH) was developed, particularly using a smartphone for transduction and quantification of optical signals. The PQQ-GDH enzyme was immobilized on SiO2 nanoparticles deposited on glass microfiber filter paper, providing a high load of the biocatalytic enzyme. The platform was tested and optimized for glucose determination using a wild type of the PQQ-GDH enzyme. The analysis was based on the fluorescence generated by the reduced form of phenazine methosulfate produced stoichiometrically to the glucose concentration. The fluorescent signals were generated at separate analytical spots on the paper support under wavelength (365 nm) UV excitation. The images of the analytical spots, dependent on the glucose concentration, were obtained using a photo camera of a standard smartphone. Then, the images were processed and quantified using software installed in a smartphone. The developed biocatalytic platform is the first step to assembling a large variety of biosensors using the same platform functionalized with artificial allosteric chimeric PQQ-dependent glucose dehydrogenase activated with different analytes. The future combination of the artificial enzymes, the presently developed analytical platform, and signal processing with a smartphone will lead to novel point-of-care and end-user biosensors applicable to virtually all possible analytes.
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Funding
This work was supported in part by the Human Frontier Science Program, project grant RGP0002/2018 to OS and EK, and by the US Department of Defense award W81XWH2010708 to EK. This paper is dedicated to the memory of Prof. Artem Melman (deceased in 2021) who was one of the leading scientists working on this project, as well as on many other research projects.
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This paper is dedicated to the memory of Artem Melman.
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Wells, P.K., Melman, A., Katz, E. et al. Fluorescent sensor based on pyrroloquinoline quinone (PQQ)-glucose dehydrogenase for glucose detection with smartphone-adapted signal analysis. Microchim Acta 189, 371 (2022). https://doi.org/10.1007/s00604-022-05466-4
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DOI: https://doi.org/10.1007/s00604-022-05466-4