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Influence of a Reduced CO2 Environment on the Secretion Yield, Potency and N-Glycan Structures of Recombinant Thyrotropin from CHO Cells

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Abstract

A consistent increase of ∼60% in the secretion yield of CHO-derived hTSH was observed by changing cell culture CO2 conditions from 5% CO2 to an air environment. The overall quality of the products obtained under both conditions was evaluated in comparison with a well-known biopharmaceutical (Thyrogen®). The N-glycans identified were of the complex type, presenting di-, tri- and tetra-antennary structures, sometimes fucosylated, 86–88% of the identified structures being sialylated at variable levels. The three most abundant structures were monosialylated glycans, representing ∼69% of all identified forms in the three preparations. The main difference was found in terms of antennarity, with 8–10% more di-antennary structures obtained in the absence of CO2 and 7–9% more tri-antennary structures in its presence. No remarkable difference in charge isomers was observed between the three preparations, the isoelectric focusing profiles showing six distinct bands in the 5.39–7.35 pI range. A considerably different distribution, with more forms in the acidic region, was observed, however, for two native pituitary preparations. All recombinant preparations showed a higher in vivo bioactivity when compared to native hTSH. Different production processes apparently do not greatly affect N-glycan structures, charge isomer distribution or bioactivity of CHO-derived hTSH.

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Acknowledgments

This work was supported by FAPESP, São Paulo, Brazil (03/11023-0 and 04/08904-7) and by the National Research Council (CNPq), Brasilia, Brazil (PQ 305108/2005-0 and PQ 301103/2006-2).

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Authors and Affiliations

  1. Biotechnology Department, IPEN-CNEN, Av. Prof. Lineu Prestes, 2242, Cidade Universitária, Sao Paulo, 05508-900, Brazil

    João Ezequiel Oliveira, Renata Damiani, Paolo Bartolini & Maria Teresa C. P. Ribela

  2. Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Vienna, Austria

    Karola Vorauer-Uhl

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  1. João Ezequiel Oliveira
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  2. Renata Damiani
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  5. Maria Teresa C. P. Ribela
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Correspondence to Maria Teresa C. P. Ribela.

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Oliveira, J.E., Damiani, R., Vorauer-Uhl, K. et al. Influence of a Reduced CO2 Environment on the Secretion Yield, Potency and N-Glycan Structures of Recombinant Thyrotropin from CHO Cells. Mol Biotechnol 39, 159–166 (2008). https://doi.org/10.1007/s12033-008-9047-6

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