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RADEs-MIP-PCa: development of rapid analysis devices for monitoring of PSA and sarcosine, Prostate Cancer (PCa) biomarkers employing biomimetic polymers based on molecular impression technology

Grant number: 22/03553-0
Support Opportunities:Regular Research Grants
Duration: December 01, 2022 - November 30, 2024
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
Principal Investigator:Maria Del Pilar Taboada Sotomayor
Grantee:Maria Del Pilar Taboada Sotomayor
Host Institution: Instituto de Química (IQ). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
Associated researchers:Ademar Wong ; Sabir khan ; Sérgio Luis Felisbino

Abstract

This project proposes to develop sensors and lateral flow devices for the determination of PSA (KLK3, kallikrein-related peptidase 3) and sarcosine. To develop the devices, molecularly imprinted polymers (MIP) will be used to replace biological systems, as they are part of one of the classes of polymeric materials applied in the identification and pre-concentration of different types of molecules with high selectivity. MIPs have been applied in different areas of knowledge and more recently in the replacement of biological materials, in immunoassays and biomimetic immunosensors.For the detection of sarcosine in artificial urine samples, a fluorescent optical sensor will be developed by covering the surface of an optical fiber tip initially with quantum-dots (QD), and then the MIP using the technique RAFT or precipitation, will be polymerized in situ. Sarcosine does not fluoresce and as its concentration increases in the sample, a quenching of the fluorescence emitted by the QD is expected. Additionally, the new material will be synthesized, optimized and evaluated in its performance, both conventionally (only the MIP) and in the core@shell format, with the quantum-dots (QD) as the core.Regarding the determination of KLK3, commonly known as PSA (prostate specific antigen), lateral flow devices (LF) will be developed aiming at speed in obtaining results, high selectivity and stability, low cost and easy handling. These are analysis systems inspired by Point of Care Tests (POCT) tests, that is, for use anywhere; without the obligation to be in a laboratory. Thus, the LF assays will be based on nitrocellulose strips modified with MIP hybrids of the core@shell type, with silica core, in place of KLK3 specific antibodies. For this, the MIP, as well as the LF strips, must be optimized. Since the synthesis of MIP, really selective for macromolecules, is not a trivial task, in this project the quantum and classical computational methods of the density functional theory based on the Kohn-Sham (KS) formalism should be used and, Molecular Dynamics and molecular modeling through the use of Molecular Docking. The calculations will be done using the free software ORCA which has the DFT and GROMACS installed for the classical calculations. These studies will make it possible to obtain really selective materials for the experimental development of materials.Since the testing of materials and devices in real samples is essential to verify the feasibility of the proposed project, initially synthetic samples of urine, serum and sperm will be analyzed, they will also be placed in contact with extracts of culture media or lysates of normal (PNT-2) and tumoral (LNCaP, PC-3 and DU145) prostatic cell lines to evaluate the efficiency of the materials in relation to the adsorption of analytes (sarcosine and KLK3) and selectivity, and for that the participation of the collaborating professor is essential , as it has expertise in the area of this type of cancer. Likewise, after approval of this project, we will start the procedures to obtain approval by the Medical Ethics Committee for the use of urine and/or sperm samples from patients, from Hospital das Clínicas de Botucatu, already diagnosed with PCa and from patients with negative diagnosis of the disease. However, we emphasize that the project proposed here can be developed even without the stage involving patients, using only human tumor cell lines, without prejudice to the results obtained, since this area of research is still new and is in rapid growth, and undoubtedly has high technological potential in the area of biotechnology. (AU)

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