Development of methodologies for protein enrichment in liquid biopsies using functionalized magnetic nanoparticles.

• Grant number: 23/18374-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: July 01, 2024
• End date: March 31, 2026
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Adriana Franco Paes Leme
• Grantee: Isamara Rodrigues Barbosa
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 18/18496-6 - The role of alcohol treated-extracellular vesicles in oral cells transformation, AP.TEM

Abstract

Proteomics allows us to analyze on a large scale the global content of proteins present indifferent biological systems through their identification and quantification, in addition to their post-translational modifications (PTMs). Literature studies, including those of our group, have demonstrated that omics-based analyzes on various biofluids allow understanding changes in these biomolecules and their possible associations with physiological and pathological processes. Although the sensitivity of techniques and equipment have been implemented over the years, there is still a limitation of proteome coverage as a function of the large dynamic concentration range. Therefore, this project proposes to develop methodologies using functionalized magnetic nanoparticles (NPMs) to expand proteome coverage in terms of number of proteins identified and quantified, such as also coverage of protein sequences, minimizing the high dynamic range of protein concentrations.proteins. To achieve this, three types of NPMs, in addition to the commercial MagResyn® SAX and Sera-MagTM SpeedBeads, will be synthesized in house, functionalized with carboxyl group, phosphatidylcholine and polyethylene glycol andcharacterized by scanning electron microscopy, X-ray photoelectron spectroscopy, diffractionX-ray, zeta potential and infrared spectroscopy. In the subsequent step, the NPMs will beincubated with samples of saliva, plasma and tear fluid from healthy individuals (n=6) for evaluation of the "corona effect" and proteins identified and quantified by spectrometry-based proteomics of masses and data analyzed by different bioinformatics strategies. The results of this study will expand knowledge of complex biofluid proteomes, prospecting selected NPMsfor future steps applied to the enrichment of patient biofluid proteomes, increasing theability to select candidate proteins for markers.