A new Chinese hamster ovary cell line expressing α2,6-sialyltransferase used as universal host for the production of human-like sialylated recombinant glycoproteins |
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Authors: | Alessandra Bragonzi Gianfranco Distefano Lorraine D Buckberry Giulia Acerbis Chiara Foglieni Damien Lamotte Gabriele Campi Annie Marc Marco R Soria Nigel Jenkins Lucia Monaco |
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Institution: | 1. DIBIT, Department of Biological and Technological Research, San Raffaele Scientific Institute, via Olgettina 58, 20132 Milan, Italy;2. Department of Biological Sciences, De Montfort University, The Gateway, Leicester LE1 9BH, UK;3. Laboratoire des Sciences du Génie Chimique, CNRS-ENSIC, 1 rue Grandville, P.O. Box 451, 54001 Nancy Cedex, France |
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Abstract: | Chinese hamster ovary (CHO) cells are widely employed to produce glycosylated recombinant proteins. Our group as well as others have demonstrated that the sialylation defect of CHO cells can be corrected by transfecting the α2,6-sialyltransferase (α2,6-ST) cDNA. Glycoproteins produced by such CHO cells display both α2,6- and α2,3-linked terminal sialic acid residues, similar to human glycoproteins. Here, we have established a CHO cell line stably expressing α2,6-ST, providing a universal host for further transfections of human genes. Several relevant parameters of the universal host cell line were studied, demonstrating that the α2,6-ST transgene was stably integrated into the CHO cell genome, that transgene expression was stable in the absence of selective pressure, that the recombinant sialyltransferase was correctly localized in the Golgi and, finally, that the bioreactor growth parameters of the universal host were comparable to those of the parental cell line. A second step consisted in the stable transfection into the universal host of cDNAs for human glycoproteins of therapeutic interest, i.e. interferon-γ and the tissue inhibitor of metalloproteinases-1. Interferon-γ purified from the universal host carried 40.4% α2,6- and 59.6% α2,3-sialic acid residues and showed improved pharmacokinetics in clearance studies when compared to interferon-γ produced by normal CHO cells. |
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Keywords: | α2 6-Sialyltransferase Stable transfection Chinese hamster ovary cell Interferon-γ Glycosylation Clearance |
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