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Establishment of scalable bioprocesses for producing virus-like particles

Abstract

Vaccine production technology named virus-like particles (VLP) has been establishing itself in the commercial applications of these pharmaceutical bioproducts. These nanostructures comprising viral proteins are safer than traditional vaccines (attenuated and inactivated virus) and more immunogenic than subunits. Among the VLP production platforms, the baculovirus/insect cell (B/IC) system has a prominent place. Two viral expression strategies to generate complex VLP are available for the B/IC: infection with multiple monocistronic baculoviruses (coinfection) and infection with a single polycistronic baculovirus (coexpression). The VLP by the B/IC are usually produced in stirred tank bioreactor (STB), to later be purified mainly depending on the scale, through density gradient centrifugation (laboratory scale) or chromatographic techniques (commercial scale). In each of these steps, multiple operational parameters must be optimized initially on a bench scale in equipment like those used on a commercial scale, in order to obtain bioprocesses that can be scaled up to the industrial implementation. Thus, the objective of this proposal will be to optimize the agitation and aeration conditions, operating mode (batch, fed-batch) in STB, as well as the chromatographic conditions (buffer system, sample volume/column volume, linear velocity) in the propagation processes of recombinant baculoviruses and in the production of VLP, using as models, the rabies virus and SARS-CoV-2, etiological agents of rabies (a neglected disease) and COVID-19, respectively. Finally, the purified and characterized VLPs will be immunologically evaluated by in vitro techniques so that the use of laboratory animals is reduced, and the proposed bioprocesses can be validated. If these goals are successfully achieved, robust bioprocesses can be transferred to later stages of development with optimized yields and productivity, as well as associated products with preliminary proven biological activity. The results of this proposal would also allow the implementation of a scalable VLP production platform for different known and unknown viral diseases. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
GUARDALINI, LUIS GIOVANI OLIVEIRA; DIAS, VINICIUS ARAGAO TEJO; LEME, JACI; BERNARDINO, THAISSA CONSONI; ASTRAY, RENATO MANCINI; DA SILVEIRA, SUELLEN REGINA; HO, PAULO LEE; TONSO, ALDO; JORGE, SORAIA ATTIE CALIL; NUNEZ, EUTIMIO GUSTAVO FERNANDEZ. Comparison of chemometric models using Raman spectroscopy for offline biochemical monitoring throughout the VLP-making upstream process. Biochemical Engineering Journal, v. 198, p. 12-pg., . (18/10538-1, 22/02713-3, 22/04711-8, 16/22780-6)

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