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Control of redox potential in hybridoma cultures: effects on MAb production, metabolism, and apoptosis

  • Fermentation, Cell Culture and Bioengineering
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Culture redox potential (CRP) has proven to be a valuable monitoring tool in several areas of biotechnology; however, it has been scarcely used in animal cell culture. In this work, a proportional feedback control was employed, for the first time, to maintain the CRP at different constant values in hybridoma batch cultures for production of a monoclonal antibody (MAb). Reducing and oxidant conditions, in the range of −130 and +70 mV, were maintained in 1-l bioreactors through automatic control of the inlet gas composition. Cultures at constant DOT, in the range of 3 and 300 %, were used for comparison. The effect of constant CRP on cell concentration, MAb production, metabolism of glucose, glutamine, thiols, oxygen consumption, and programmed cell death, was evaluated. Reducing conditions resulted in the highest viable cell and MAb concentrations and thiols production, whereas specific glucose and glutamine consumption rates remained at the lowest values. In such conditions, programmed cell death, particularly apoptosis, occurred only after nutrient exhaustion. The optimum specific MAb production rate occurred at intermediate CRP levels. Oxidant conditions resulted in a detrimental effect in all culture parameters, increasing the specific glucose, glutamine, and oxygen consumption rates and inducing the apoptotic process, which was detected as early as 24 h even when glutamine and glucose were present at non-limiting concentrations. In most cases, such results were similar to those obtained in control cultures at constant DOT.

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Acknowledgments

A. Meneses-Acosta thanks the Universidad Autónoma del Estado de Morelos for the financial support to develop this work during the sabbatical year.

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Authors and Affiliations

  1. Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal. 510-3, 62250, Cuernavaca, Morelos, Mexico

    Angélica Meneses-Acosta, Alfonso Gómez & Octavio T. Ramírez

  2. Facultad de Farmacia, Universidad Autónoma del Estado de Morelos, Av. Universidad 1010, Col. Chamilpa, 62100, Cuernavaca, Morelos, Mexico

    Angélica Meneses-Acosta

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  1. Angélica Meneses-Acosta
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  2. Alfonso Gómez
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  3. Octavio T. Ramírez
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Correspondence to Octavio T. Ramírez.

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Meneses-Acosta, A., Gómez, A. & Ramírez, O.T. Control of redox potential in hybridoma cultures: effects on MAb production, metabolism, and apoptosis. J Ind Microbiol Biotechnol 39, 1189–1198 (2012). https://doi.org/10.1007/s10295-012-1125-x

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