Optimization of purification platforms based on chromatographic techniques to obtain rabies and SARS-CoV-2 VLP

• Grant number: 25/10314-0
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: July 01, 2025
• End date: June 30, 2029
• Field of knowledge: Health Sciences - Pharmacy - Pharmaceutical Technology
• Principal Investigator: Eutimio Gustavo Fernández Núñez
• Grantee: Milena Miyu Teruya
• 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

As objectives from the Immunization Agenda 2030 (IA2030), the World Health Organization has stated the necessity to amplify access to vaccines to all countries to balance its distribution and raise vaccination coverage, as these represent the main prophylaxis method for most infectious diseases. Vaccines against viral diseases caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), related to the last pandemic episode, with millions of cases and deaths, and by rabies virus, a lethal disease with thousands of deaths annually, are among the vaccines included on the IA2030 implementation list. Amidst vaccine platforms, use of virus-like particles (VLP) is under study, since these are non-infectious, induce a strong immune response, and are fast to produce. Nevertheless, to remove process-related contaminants, like cell debris, genetic material, proteins, and others, purification steps are needed to obtain the VLP. Low-pressure liquid chromatography (LPLC) is a usual purification method to obtain pharmaceutical proteins, which can be performed using different types of resins, such as ionic exchange, hydrophobic interactions, and others. Therefore, this project aims to optimize rabies and SARS-CoV-2 VLP purification parameters through two LPLC downstream platforms. It is expected to establish optimal buffer solution for each resin and chromatographic step, establish working variables such as sample volume and mobile phase linear velocity, and finally, define the best operational conditions for purification of these VLP according to product purity and yield obtained with these methods.