Chemometrics and analytical blank on the at-line monitoring of Zika-VLP production using near-infrared spectroscopy

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Rabello, Julia Publio ; Cavalcante, Paulo Eduardo da Silva ; Leme, Jaci ; Dias, Vinicius Aragao Tejo ; Barrence, Fernanda Angela Correia ; Guardalini, Luis Giovani de Oliveira ; Bernardino, Thaissa Consoni ; Nunes, Robson ; Barros, Iago Henrique ; Tonso, Aldo ; Jorge, Soraia Attie Calil ; Nunez, Eutimio Gustavo Fernandez

• Tipo de documento: Artigo Científico
• Fonte: SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY; v. 326, p. 12-pg., 2024-10-05.

Resumo

The Zika disease caused by the Zika virus was declared a Public Health Emergency by the World Health Union (WHO), with microcephaly as the most critical consequence. Aiming to reduce the spread of the virus, biopharmaceutical organizations invest in vaccine research and production, based on multiple platforms. A crescent vaccine production approach is based on virus-like particles (VLP), for not having genetic material in its composition, hypoallergenic and non-mutant character. For bioprocess, it is essential to have means of real-time monitoring, which can be assessed using process analysis techniques such as Near-infrared (NIR) spectroscopy, that can be combined with chemometric methods, like Partial-Least Squares (PLS) and Artificial Neural Networks (ANN) for prediction of biochemical variables. This work proposes a biochemical Zika VLP upstream production at-line monitoring model using NIR spectroscopy comparing sampling conditions (with or without cells), analytical blank (air, ultrapure water), and spectra pre-processing approaches. Seven experiments in a benchtop bioreactor using recombinant baculovirus/Sf9 Sf9 insect cell platform in serum-free medium were performed to obtain biochemical and spectral data for chemometrics modeling (PLS and ANN), composed by a random data split (80 % calibration, 20 % validation) for cross-validation of the PLS models and 70 % training, 15 % testing, 15 % validation for ANN. The best models generated in the present work presented an average absolute error of 1.59 x 105 5 cell/mL for density of viable cells, 2.37% for cell viability, 0.25 g/L for glucose, 0.007 g/L for lactate, 0.138 g/L for glutamine, 0.18 g/L for glutamate, 0,003 g/L for ammonium, and 0.014 g/L for potassium. (AU)

• Processo FAPESP: 23/09463-5 - Comprimento de onda do laser e condicionamento da amostra no monitoramento bioquímico por espectroscopia Raman da produção de VLP de SARS-CoV-2
• Beneficiário: Júlia Públio Rabello
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 22/02713-3 - Estabelecimento de bioprocessos escalonáveis para produção de partículas semelhantes a vírus
• Beneficiário: Eutimio Gustavo Fernández Núñez
• Modalidade de apoio: Auxílio à Pesquisa - Projeto Inicial