Applying the Quality-by-design approach to optimize the upstream stage of Rabies VLP production

• Grant number: 24/22430-1
• Support Opportunities: Scholarships in Brazil - Master
• Start date: July 01, 2025
• End date: June 30, 2027
• Field of knowledge: Health Sciences - Pharmacy - Pharmaceutical Technology
• Principal Investigator: Eutimio Gustavo Fernández Núñez
• Grantee: Júlia Públio Rabello
• Host Institution: Escola de Artes, Ciências e Humanidades (EACH). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 22/02713-3 - Establishment of scalable bioprocesses for producing virus-like particles, AP.PNGP.PI

Abstract

Rabies virus (RABV), belonging to the Lyssavirus genus, is the causative agent of rabies, a zoonotic disease with a fatality rate close to 100%, with more than 74,000 victims annually. Transmitted mainly by dogs, rabies affects the central nervous system and is fatal in humans, highlighting the need for prophylaxis. Despite the efficacy of existing vaccines, their production is expensive and not easily accessible for endemic regions, reinforcing the search for innovative alternatives, such as vaccines based on virus-like particles (VLP). VLP mimics the structure of native viruses without containing genetic material, ensuring safety and high immunogenicity. The production of VLP depends on factors such as culture conditions, specific parameters such as time of infection (TOI), dissolved oxygen (DO), and the chosen expression system. The Baculovirus-insect cell system is widely used in the production of biopharmaceutical proteins and VLP due to its ability to perform post-translational modifications, high protein production, and easy scalability. Quality by Design (QbD) is an approach suggested by drug regulatory agencies to optimize bioprocesses, which allows identifying and controlling critical variables to ensure product quality and the technical-economic viability of the process. Thus, this study proposes to evaluate, at the bench level, the ideal conditions to produce rabies VLP, considering parameters such as cell host/culture medium, TOI, harvest time (HT) and OD through statistical experimental designs. The aim is to develop an efficient and scalable process to enhance global access to rabies immunization, especially in resource-limited regions.