Biochemical monitoring throughout all stages of rabies virus-like particles production by Raman spectroscopy using global models

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Guardalini, Luis Giovani Oliveira ; Cavalcante, Paulo Eduardo da Silva ; Leme, Jaci ; de Mello, Renata Gois ; Bernardino, Thaissa Consoni ; Astray, Renato Mancini ; Barbosa, Eduardo ; da Silveira, Suellen Regina ; Ho, Paulo Lee ; Tonso, Aldo ; Jorge, Soraia Attie Calil ; Nunez, Eutimio Gustavo Fernandez

• Document type: Journal article
• Source: Journal of Biotechnology; v. 363, p. 13-pg., 2023-01-06.

Abstract

This work aimed to quantify growth and biochemical parameters (viable cell density, Xv; cell viability, CV; glucose, lactate, glutamine, glutamate, ammonium, and potassium concentrations) in upstream stages to obtain rabies virus-like particles (rabies VLP) from insect cell-baculovirus system using on-line and off-line Raman spectra to calibrate global models with minimal experimental data. Five cultivations in bioreactor were per-formed. The first one comprised the growth of uninfected Spodoptera frugiperda (Sf9) cells, the second and third runs to obtain recombinant baculovirus (rBV) bearing Rabies G glycoprotein and matrix protein, respectively. The fourth one involved the generation of rabies VLP from rBVs and the last one was a repetition of the third one with cell inoculum infected by rBV. The spectra were acquired through a Raman spectrometer with a 785-nm laser source. The fitted Partial Least Square models for nutrients and metabolites were comparable with those previously reported for mammalian cell lines (Relative error < 15 %). However, the use of this chemometrics approach for Xv and CV was not as accurate as it was for other parameters. The findings from this work established the basis for bioprocess Raman spectroscopical monitoring using insect cells for VLP manufacturing, which are gaining ground in the pharmaceutical industry. (AU)

• FAPESP's process: 16/22780-6 - Arboviruses VLP production in insect cells.
• Grantee: Soraia Attie Calil Jorge
• Support Opportunities: Regular Research Grants
• FAPESP's process: 18/10538-1 - Characterization of recombinant antigens and immune response of a new rabies vaccine candidate
• Grantee: Renato Mancini Astray
• Support Opportunities: Regular Research Grants