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Human Cells as Platform to Produce Gamma-Carboxylated Proteins

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Recombinant Glycoprotein Production

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1674))

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Abstract

The gamma-carboxylated proteins belong to a family of proteins that depend on vitamin K for normal biosynthesis. The major representative gamma-carboxylated proteins are the coagulation system proteins, for example, factor VII, factor IX, factor X, prothrombin, and proteins C, S, and Z. These molecules have harbored posttranslational modifications, such as glycosylation and gamma-carboxylation, and for this reason they need to be produced in mammalian cell lines. Human cells lines have emerged as the most promising alternative to the production of gamma-carboxylated proteins. In this chapter, the methods to generate human cells as a platform to produce gamma-carboxylated proteins, for example the coagulation factors VII and IX, are presented. From the cell line modification up to the vitamin K adaptation of the produced cells is described in the protocols presented in this chapter.

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Acknowledgments

The authors acknowledge São Paulo Research Foundation—FAPESP (2015/19017-6), Conselho Nacional de Pesquisa—CNPq (142406/2016-3), Centro de Pesquisa, Inovação e Difusão (CEPID), and National Institute of Science and Technology in Stem Cell and Cell Therapy—INCTC for financial support and Sandra Navarro for drawing the figures.

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Authors and Affiliations

  1. Center for Cell-based Therapy CTC, Ribeirão Preto Medical School, University of São Paulo, Av. Tenente Catão Roxo 2501, Monte Alegre, 14051-140, Ribeirão Preto, SP, Brazil

    Aline de Sousa Bomfim, Marcela Cristina Corrêa de Freitas, Dimas Tadeu Covas & Elisa Maria de Sousa Russo

  2. School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Av. Tenente Catão Roxo 2501, Monte Alegre, 14051-140, Ribeirão Preto, SP, Brazil

    Aline de Sousa Bomfim & Elisa Maria de Sousa Russo

  3. Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Av. Tenente Catão Roxo 2501, Monte Alegre, 14051-140, Ribeirão Preto, SP, Brazil

    Dimas Tadeu Covas

Authors
  1. Aline de Sousa Bomfim
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  2. Marcela Cristina Corrêa de Freitas
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  3. Dimas Tadeu Covas
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  4. Elisa Maria de Sousa Russo
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Corresponding author

Correspondence to Aline de Sousa Bomfim .

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Editors and Affiliations

  1. Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, Brazil

    Virgínia Picanço-Castro

  2. School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, Brazil

    Kamilla Swiech

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de Sousa Bomfim, A., de Freitas, M.C.C., Covas, D.T., de Sousa Russo, E.M. (2018). Human Cells as Platform to Produce Gamma-Carboxylated Proteins. In: Picanço-Castro, V., Swiech, K. (eds) Recombinant Glycoprotein Production. Methods in Molecular Biology, vol 1674. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7312-5_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7312-5_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7311-8

  • Online ISBN: 978-1-4939-7312-5

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