1. Biochemical monitoring throughout all stages of rabies virus-like particles production by Raman spectroscopy using global models.
- Author
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Oliveira Guardalini, Luis Giovani, da Silva Cavalcante, Paulo Eduardo, Leme, Jaci, Gois de Mello, Renata, Consoni Bernardino, Thaissa, Mancini Astray, Renato, Barbosa, Eduardo, da Silveira, Suellen Regina, Lee Ho, Paulo, Tonso, Aldo, Attie Calil Jorge, Soraia, and Fernández Núñez, Eutimio Gustavo
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VIRUS-like particles , *GLUTAMINE , *RABIES , *RAMAN spectroscopy , *FALL armyworm , *EXTRACELLULAR matrix proteins , *MANUFACTURING cells , *GLUTAMINE synthetase - 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 performed. 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. [Display omitted] • Modeling from Raman spectra using PLS for rabies VLP production monitoring. • Raman spectral range pick for monitoring biochemically rabies VLP process. • Effective online biochemical monitoring over rabies VLP production with Raman data. • Suitable biochemical monitoring over rabies VLP production with offline Raman data. • Assessed parameters relative error was less than 15 % using online or offline data. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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