Enzyme Array: A New Approach for Studying Metabolism

• Grant number: 25/10680-6
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: April 01, 2026
• End date: December 31, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Metabolism and Bioenergetics
• Principal Investigator: Calina Liu Yu Ying
• Grantee: Calina Liu Yu Ying
• Principal investigators: João Victor Virgilio da Silva ; Larissa Menezes dos Reis

Abstract

Metabolism is a vast field of study that relies on high-cost, complex methodologies requiring specific equipment and intricate analyses, often challenging to interpret. As an alternative strategy, many research groups use tools that are not specifically designed for metabolic questions, frequently obtaining results that may not accurately reflect the biological process being evaluated. Given the limitations observed in studying metabolism, we propose a strategy based on enzymatic activity assays, enabling the evaluation of multiple enzymes within the same experiment-what we call the "enzyme array". Our differentiator will be the option for researchers to customize a 96- or 384-well plate with assays for various enzymes within the same metabolic pathway (to assess regulatory points of their pathway of interest) or with activity assays for enzymes involved in different biological processes (allowing preliminary insights to support investment in more costly techniques, such as metabolomics with labeled nutrients). Initially, our target audience will be researchers focused on metabolism or those working in various fields but with specific questions related to metabolic pathways. Our primary goal is to offer an affordable, routine-use product with high reproducibility and ease of implementation across different labs. To achieve this, during the PIPE Phase I project, we will finalize the optimization of various enzymatic activity assays (targeting metabolic pathways: glycolysis, TCA cycle anaplerosis, TCA cycle function, mitochondrial complexes, cellular redox balance) across different cell types (MDA-MB-231, B16F10, HEK 293, NIH 3T3, RAW 2647, and Jurkat lines). We will also evaluate cost-effective kit assembly methods by reducing plastic materials and shipping costs. Finally, we will develop a user-friendly analysis method, enabling a robust and comprehensive interpretation of results. (AU)