Journal
ACS APPLIED MATERIALS & INTERFACES
Volume 13, Issue 22, Pages 25624-25634Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsami.1c02505
Keywords
alpha-glucosidase inhibitors; nanobiosensor; fluorescent conjugated polymers; liposomal nanoparticles; acarbose; multiwell plate; antidiabetic drugs
Funding
- Spanish Ministry of Economy, Industry, and Competitiveness [MAT-2017-86805-R]
- Generalitat Valenciana - Conselleria d'Educacio Investigacio Cultura i Esport
- EU-FEDER Una forma de hacer Europa [GVA-IDIFEDER 2018/020]
- Ministry of Science and Innovation
- EU-FEDER [EQC2019-005842-P]
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A stable and reusable fluorescent multisample nano-biosensor has been developed for detecting alpha-glucosidase inhibitors. It can perform multiple measurements in samples containing different inhibitors and is useful for both drug screening and quantification.
A highly stable and reusable fluorescent multisample nano-biosensor for the detection of alpha-glucosidase inhibitors has been developed by coupling fluorescent liposomal nanoparticles based on conjugated polymers (L-CPNs) to the enzyme alpha-glucosidase, one of the main target enzymes in the treatment of type 2 diabetes. The mechanism of sensing is based on the fluorescence turn-on of L-CPNs by p-nitrophenol (PNP), the end product of the enzymatic hydrolysis of p-nitrophenyl-alpha-D-glucopyranoside. L-CPNs, composed of lipid vesicles coated with a blue-emitting cationic polyfluorene, were designed and characterized to obtain a good response to PNP. Two nanobiosensor configurations were developed in this study. In the first step, a single-sample nanobiosensor composed of L-CPNs and alpha-glucosidase entrapped in a sol-gel glass was developed in order to characterize and optimize the device. In the second part, the nanobiosensor was integrated and adapted to a multiwell microplate and the possibility of reusing it and performing multiple measurements simultaneously with samples containing different alpha-glucosidase inhibitors was investigated. Using super-resolution confocal microscopy, L-CPNs could be visualized within the sol-gel matrix, and the quenching of their fluorescence, induced by the substrate, was directly observed in situ. The device was also shown to be useful not only as a platform for screening of antidiabetic drugs but also for quantifying their presence. The latter application was successfully tested with the currently available drug, acarbose.
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