A multivariate calibration procedure for UV/VIS spectrometric monitoring of BHK-21 cell metabolism and growth

Authors

  • Jaci Leme,

    1. Laboratório Especial de Pesquisa e Desenvolvimento em Imunológicos Veterinários, Instituto Butantan. Av. Vital Brasil, 1500, São Paulo, SP–Brazil
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    • Both authors contributed equally to this work.

  • Eutimio Gustavo Fernández Núñez,

    Corresponding author
    1. Dept. de Engenharia Química, Laboratório de Células Animais, Escola Politécnica, Universidade de São Paulo. Av. Prof. Luciano Gualberto, São Paulo, SP–Brazil
    2. Dept. de Ciências Biológicas, Universidade Estadual Paulista “Júlio de Mesquita Filho” Campus-Assis, Avenida Dom Antonio 2100, Bairro Parque Universitário, Assis, SP – Brasil
    • Correspondence concerning this article should be addressed to E. G. Fernández Núñez at eutimiocu@yahoo.com.

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    • Both authors contributed equally to this work.

  • Letícia de Almeida Parizotto,

    1. Dept. de Engenharia Química, Laboratório de Células Animais, Escola Politécnica, Universidade de São Paulo. Av. Prof. Luciano Gualberto, São Paulo, SP–Brazil
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  • Wagner Antonio Chagas,

    1. Laboratório Especial de Pesquisa e Desenvolvimento em Imunológicos Veterinários, Instituto Butantan. Av. Vital Brasil, 1500, São Paulo, SP–Brazil
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  • Erica Salla dos Santos,

    1. Laboratório Especial de Pesquisa e Desenvolvimento em Imunológicos Veterinários, Instituto Butantan. Av. Vital Brasil, 1500, São Paulo, SP–Brazil
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  • Aline Tojeira Prestia Caricati,

    1. Laboratório Especial de Pesquisa e Desenvolvimento em Imunológicos Veterinários, Instituto Butantan. Av. Vital Brasil, 1500, São Paulo, SP–Brazil
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  • Alexandre Gonçalves de Rezende,

    1. Laboratório de Imunologia Viral, Instituto Butantan. Av. Vital Brasil, Butantã, São Paulo, SP–Brazil
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  • Bruno Labate Vale da Costa,

    1. Dept. de Engenharia Química, Laboratório de Células Animais, Escola Politécnica, Universidade de São Paulo. Av. Prof. Luciano Gualberto, São Paulo, SP–Brazil
    2. Laboratório de Biotecnologia Industrial, Núcleo de Bionanomanufatura, Instituto de Pesquisas Tecnológicas do Estado de São Paulo, Av. Prof. Almeida Prado 532 Cid. Universitária - Butantã, São Paulo, SP–Brazil
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  • Daniela Cristina Ventini Monteiro,

    1. Laboratório de Imunologia Viral, Instituto Butantan. Av. Vital Brasil, Butantã, São Paulo, SP–Brazil
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  • Vera Lucia Lopes Boldorini,

    1. Laboratório de Imunologia Viral, Instituto Butantan. Av. Vital Brasil, Butantã, São Paulo, SP–Brazil
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  • Soraia Attie Calil Jorge,

    1. Laboratório de Imunologia Viral, Instituto Butantan. Av. Vital Brasil, Butantã, São Paulo, SP–Brazil
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  • Renato Mancini Astray,

    1. Laboratório de Imunologia Viral, Instituto Butantan. Av. Vital Brasil, Butantã, São Paulo, SP–Brazil
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  • Carlos Augusto Pereira,

    1. Dept. de Engenharia Química, Laboratório de Células Animais, Escola Politécnica, Universidade de São Paulo. Av. Prof. Luciano Gualberto, São Paulo, SP–Brazil
    2. Laboratório de Imunologia Viral, Instituto Butantan. Av. Vital Brasil, Butantã, São Paulo, SP–Brazil
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  • Celso Pereira Caricati,

    1. Laboratório Especial de Pesquisa e Desenvolvimento em Imunológicos Veterinários, Instituto Butantan. Av. Vital Brasil, 1500, São Paulo, SP–Brazil
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  • Aldo Tonso

    1. Dept. de Engenharia Química, Laboratório de Células Animais, Escola Politécnica, Universidade de São Paulo. Av. Prof. Luciano Gualberto, São Paulo, SP–Brazil
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Abstract

Monitoring mammalian cell culture with UV–vis spectroscopy has not been widely explored. The aim of this work was to calibrate Partial Least Squares (PLS) models from off-line UV–vis spectral data in order to predict some nutrients and metabolites, as well as viable cell concentrations for mammalian cell bioprocess using phenol red in culture medium. The BHK-21 cell line was used as a mammalian cell model. Spectra of samples taken from batches performed at different dissolved oxygen concentrations (10, 30, 50, and 70% air saturation), in two bioreactor configurations and with two strategies to control pH were used to calibrate and validate PLS models. Glutamine, glutamate, glucose, and lactate concentrations were suitably predicted by means of this strategy. Especially for glutamine and glucose concentrations, the prediction error averages were lower than 0.50 ± 0.10 mM and 2.21 ± 0.16 mM, respectively. These values are comparable with those previously reported using near infrared and Raman spectroscopy in conjunction with PLS. However, viable cell concentration models need to be improved. The present work allows for UV–vis at-line sensor development, decrease cost related to nutrients and metabolite quantifications and establishment of fed-batch feeding schemes. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:241–248, 2014

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