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Na-butyrate increases the production and α2,6-sialylation of recombinant interferon-γ expressed by α2,6- sialyltransferase engineered CHO cells

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Abstract

A non-human like glycosylation pattern in human recombinant glycoproteins expressed by animal cells may compromise their use as therapeutic drugs. In order to correct the CHO glycosylation machinery, a CHO cell line producing recombinant human interferon- γ (IFN) was transformed to replace the endogenous pseudogene with a functional copy of the enzyme α2,6-sialyltransferase (α2,6-ST). Both the parental and the modified CHO cell line were propagated in serum-free batch culture with or without 1 mM sodium butyrate. Although Na-butyrate inhibited cell growth, IFN concentration was increased twofold. The IFN sialylation status was determined using linkage specific sialidases and HPLC. Under non- induced conditions, IFN expressed by α2,6-engineered cells contained 68% of the total sialic acids in the α2,6- conformation and the overall molar ratio of sialic acids to IFN was 2.3. Sodium butyrate addition increased twofold the molar ratio of total sialic acids to IFN and 82% of total sialic acids on IFN were in the α2,6-conformation. In contrast, no effect of the sodium butyrate was noticed on the sialylation of the IFN secreted by the α2,6-ST deficient parental cell line. This study deals for the first time with the effect of Na-butyrate on CHO cells engineered to produce human like sialylation.

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Authors and Affiliations

  1. Laboratoire des Sciences du Génie Chimique, CNRS-ENSIC, 1, rue Grandville, BP 451, 54001, Nancy Cedex, France

    Damien Lamotte, Jean-Marc Engasser & Annie Marc

  2. Department of Biological Sciences, De Montfort University, Leicester, LEI 9BH, UK

    Lorraine Buckberry & Nigel Jenkins

  3. San Raffaele Scientific Institute, DIBIT, via Olgettina 60, 20132, Milano, Italy

    Lucia Monaco & Marco Soria

Authors
  1. Damien Lamotte
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  2. Lorraine Buckberry
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  3. Lucia Monaco
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  4. Marco Soria
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  5. Nigel Jenkins
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  6. Jean-Marc Engasser
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  7. Annie Marc
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Lamotte, D., Buckberry, L., Monaco, L. et al. Na-butyrate increases the production and α2,6-sialylation of recombinant interferon-γ expressed by α2,6- sialyltransferase engineered CHO cells. Cytotechnology 29, 55–64 (1999). https://doi.org/10.1023/A:1008080432681

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