Project EMU: Acquisition of equipment that automates the separation of proteins by size and immunodetection in traditional Western blotting by chemiluminescence or fluorescence - JESS from Protein Simple

• Grant number: 23/17365-3
• Support Opportunities: Research Infrastructure Program - Small Medium Sized
• Start date: September 01, 2024
• End date: August 31, 2027
• Field of knowledge: Agronomical Sciences - Food Science and Technology
• Principal Investigator: Elizabeth Aparecida Ferraz da Silva Torres
• Grantee: Elizabeth Aparecida Ferraz da Silva Torres
• Host Institution: Faculdade de Saúde Pública (FSP). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Marcelo Macedo Rogero ; Regina Mara Fisberg

Abstract

The Western dietary pattern promotes an increased prevalence of overweight and obesity, accompanied by physiological disorders such as hyperglycemia, hyperinsulinemia, hyperlipidemia, and hypertension, which, in turn, induce oxidative stress and an elevation of pro-inflammatory mediators. These conditions contribute to the development of insulin resistance and endothelial dysfunction. These metabolic alterations predispose individuals to the development of metabolic syndrome, and the resulting excess body fat can lead to chronic low-grade inflammation, where immune system activation occurs in the absence of infection or autoimmune disease, affecting not only a specific region or tissue but various physiological systems.Therefore, it is of paramount importance to investigate proteins related to these systems and their interaction with dietary components and complex matrices. The implementation of the JESS system aims to provide high-quality analytical solutions for research related to specific proteins and their relationship with diseases and individual health. This innovative technology combines ELISA techniques (absolute quantification of specific proteins) and Western Blotting (relative quantification of specific proteins with imaging) into a single analysis, offering speed and the use of minimal volumes of reagents, samples, and antibodies.The automation brought by the JESS equipment eliminates many error-prone steps associated with Western Blotting and ELISA techniques, as adding samples to specific microplates is sufficient to eliminate complex washing, incubation, and detection steps. Typically, Western Blotting analyses take about 2 to 3 days, and ELISA analyses take about 4 to 8 hours. JESS analysis lasts approximately 3 hours, reducing sample, reagent, and antibody volumes by up to 90% compared to traditional techniques. Furthermore, the equipment requires no specific preparation for installation-a one-meter by fifty-centimeter bench and a power outlet are sufficient-and it does not require short or medium-term maintenance. The Nutrition Department has the technical capacity and infrastructure necessary for the acquisition of the equipment, which will be widely used by researchers throughout Brazil.Keywords: JESS, Protein Simple, protein identification and quantification. (AU)