"Improvement and Scaling up of Cell-free Protein Synthesis, SynthEasy® Kits"

• Grant number: 24/04215-6
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: June 01, 2024
• End date: May 31, 2026
• Field of knowledge: Interdisciplinary Subjects
• Principal Investigator: Phelipe Augusto Mariano Vitale
• Grantee: Phelipe Augusto Mariano Vitale
• Company: Biolinker Biologia Sintética Eireli
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais
• City: Cotia
• Pesquisadores principais: Douglas Dalle Luche
• Associated research grant: 23/09974-0 - "Improvement and Scaling up of Cell-free Protein Synthesis, SynthEasy® Kits", AP.PIPE
• Associated scholarship(s): 24/17245-0 - Improvement and scaling up of cell-free protein synthesis, SynthEasy® kits, BP.PIPE

Abstract

Biolinker is the only domestic company to market a biotechnology tool called Cell-Free Protein Synthesis (CFPS), which enables the accelerated production of proteins of scientific and commercial interest. This technology, whose kits are based on extracts of Escherichia coli, overcomes technical challenges present in production within living organisms and has already received funding from FAPESP for the development of a lyophilized version (2019/16625-6). CFPS has proven fundamental for biotechnological processes that require reproducibility and stability of protein production levels, linearity, and scalability.During the development phase of the lyophilized version, the kits underwent several modifications in their composition to maintain the yield observed before lyophilization and demonstrate an increased shelf-life in months. However, there are still challenges to be overcome such as high variability between batches and difficulty in obtaining significant quantities of complex proteins such as antibodies, which are targets of biotechnological interest.The next step in the development of the CFPS kit involves research for scaling up the process, given that there is significant variation in the product quality between the batches produced. The use of Nuclear Magnetic Resonance (NMR) technique presents itself as a crucial step to analyze the extracts obtained, correlate the presence of certain molecules throughout the protein expression process and thus identify the cause of variability between batches. Therefore, the goal of this study is to establish the best production process for the CFPS kits.During the development phase of the lyophilized version, the kits underwent several modifications in composition, were subjected to lyophilization, managed to maintain the yield of protein expression, and had their shelf life extended. However, there are still challenges to be overcome, such as high variability between batches and difficulty in obtaining significant quantities of complex proteins, which are targets of biotechnological interest.The next stage in the development of the CFPS kit involves research for the scaling of the process, given that there is significant variation in the quality of the product between batches produced. The use of the Nuclear Magnetic Resonance (NMR) technique for small molecules is a crucial step to analyze the obtained extracts, correlate the presence of certain molecules throughout the protein expression process, and thus identify the cause of variability between batches. Therefore, the ultimate goal of this study is to establish the best production process for the CFPS kits. (AU)