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Article

New Label-Free Biosensing for the Evaluation of the AX-024 Inhibitor: Case Study for the Development of New Drugs in Autoimmune Diseases

1
Center of Biomedical Technology, Optics, Photonics and Biophotonics Laboratory, Universidad Politécnica de Madrid, Campus Montegancedo, Pozuelo de Alarcón, 28223 Madrid, Spain
2
BioOptical Detection SL, Centro de Empresas, Campus Montegancedo, 28223 Madrid, Spain
3
Department of Applied Physics and Materials Engineering, Escuela Técnica Superior de Ingenieros Industriales, Universidad Politécnica de Madrid, C/ José Gutierrez Abascal 2, 28006 Madrid, Spain
*
Authors to whom correspondence should be addressed.
Academic Editor: Laura Micheli
Sensors 2022, 22(3), 1218; https://doi.org/10.3390/s22031218
Received: 27 December 2021 / Revised: 28 January 2022 / Accepted: 2 February 2022 / Published: 5 February 2022
(This article belongs to the Section Biosensors)
We developed a new label-free assay to evaluate the inhibition capacity of AX-024 by means of a new Point-of-Care (PoC) device for application in the development of new drugs in autoimmune diseases. The technology of PoC is based on interferometric optical detection method (IODM). For this purpose, we have optimized and developed an assay protocol whereby a Glutathione S-Transferase modified protein (GST-SH3.1), which contains a functional domain of a protein involved in T-cell activation, together with the AX-024 inhibitor has been studied. The chips used are a sensing surface based on nitrocellulose. We used streptavidin and a biotinylated peptide as links for the immobilization process on the sensing surface. The biotinylated peptide and AX-024 inhibitor compete for the same functional group of the GST-SH3.1 modified protein. When the inhibitor binds its binding site on GST-SH3.1, the biotinylated peptide cannot bind to its pocket on the protein. This competition reduces the total molecular mass of protein fixed onto the biosensor. In order to quantify the inhibition capacity of AX-024, several Ax-024:GST-SH3.1 ratios have been studied. We have compared the read-out signal for GST-SH3.1 protein not interfered by the drug, which served as a positive blank, and the response of the GST-SH3.1 modified protein blocked by the inhibitor. The technology has been correlated with confocal fluorescence microscopy. View Full-Text
Keywords: label-free detection; point-of-care device; AX-024 inhibitor; GST-SH3.1 protein; autoimmune disease; fluorescence versus label-free detection; confocal microscopy label-free detection; point-of-care device; AX-024 inhibitor; GST-SH3.1 protein; autoimmune disease; fluorescence versus label-free detection; confocal microscopy
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MDPI and ACS Style

Ramírez, Y.; Laguna, M.F.; Holgado, M. New Label-Free Biosensing for the Evaluation of the AX-024 Inhibitor: Case Study for the Development of New Drugs in Autoimmune Diseases. Sensors 2022, 22, 1218. https://doi.org/10.3390/s22031218

AMA Style

Ramírez Y, Laguna MF, Holgado M. New Label-Free Biosensing for the Evaluation of the AX-024 Inhibitor: Case Study for the Development of New Drugs in Autoimmune Diseases. Sensors. 2022; 22(3):1218. https://doi.org/10.3390/s22031218

Chicago/Turabian Style

Ramírez, Yolanda, María Fe Laguna, and Miguel Holgado. 2022. "New Label-Free Biosensing for the Evaluation of the AX-024 Inhibitor: Case Study for the Development of New Drugs in Autoimmune Diseases" Sensors 22, no. 3: 1218. https://doi.org/10.3390/s22031218

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