Conjugação catalisada por enzima combinada com intensificação de bioprocesso: geração de nanopartículas de ferritina funcionalizadas como vacinas

The first objective of this project is to develop a novel modular assembly method to functionalize ferritin nanoparticles. The innovation here is two-fold: i) Conjugation strategy: use of tyrosinase as enzyme catalyzing the irreversible, site-specific conjugation of target antigens (i.e. SARS-CoV-2 RBD protein and RVF virus Gn glycoprotein) to ferritin nanoparticles, resulting in predictable, stable and homogenous functionalized ferritin nanoparticles. ii) Antigen display strategy: use of ferritin nanoparticles to display target antigens in their native conformations in a highly repetitive fashion, thus stimulating a strong humoral and cellular immune response. The second objective of this project is to design a production platform capable of manufacturing high quantities of vaccine in a robust, fast and economic way. The innovation here is two-fold: i) Producer cell line identification strategy: combination of cross-platform expression screening with a multi-analytical approach to assist on the selection of best producer cell line. ii) Bioprocess intensification strategy: combination of optimized high-cell-density (HCD) perfusion cultures with integration of upstream and downstream bioprocessing to generate an high-yield, fully continuous manufacturing process. The third objective of this project is to demonstrate the efficacy of the ferritin-based vaccine candidates produced for two diseases distinguished as priority under WHO’s R&D Blueprint: COVID-19 and RVF. The proposed research addresses significant challenges in vaccine development and manufacturing. It aims to overcome several key hurdles, including the need for more efficient vaccine designs, the demand for scalable and cost-effective manufacturing methods, and the necessity for rapid response capabilities during pandemics. By focusing on developing a novel modular assembly method for functionalizing ferritin nanoparticles and designing a robust, fast, low-cost production platform capable of manufacturing vaccines in large quantities, the research directly tackles these challenges. The objectives outlined demonstrate a high level of ambition and innovation, extending beyond the current state of the art in vaccine development and manufacturing. The proposed modular assembly method for functionalizing ferritin nanoparticles, employing tyrosinase for site-specific conjugation of antigens, represents a novel approach in vaccine design. In addition, the integration of optimized high-cell-density cultures with downstream processing to establish a fully continuous manufacturing process is an ambitious endeavor that pushes the boundaries of current manufacturing practices. Overall, the research presents novel concepts and approaches that have the potential to significantly advance the field of vaccine development and manufacturing. This project will have impact at various levels, from advancing the state-of-the-art to PI’s career development: - Advance the state-of-the-art: impact on human health (prevention of disease), economy (value-added products, hospitalization savings), industrial biotechnology (new biological toolkits, engineering strategies, vaccine production platform), vaccines design (modular assembly method). - PI’s career development: widen network of research collaborations, strengthen competitiveness in the vaccines area, improve inter-personal relationship management skills, improve planning and decision-making skills, foster maturity and independence.