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Genetic engineering of recombinant glycoproteins and glycosylation pathway in mammalian host cells

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Abstract

The analysis of many natural glycoproteins and their recombinant counterparts from mammalian hosts has revealed that the basic oligosaccharide structures and the site occupancy of glycosylated polypeptides are primarily dictated by the protein conformation.

The equipment of many frequently used host cells (e.g. BHK-21 and CHO-cells) with glycosyltransferases, nucleotide-sugar synthases and transporters appears to be sufficient to guarantee complex-type glycosylation of recombinant proteins with a high degree of terminal α2-3 sialylation even under high expression conditions. Some human tissue-specific terminal carbohydrate motifs are not synthesized by these cells since they lack the proper sugar-transferring enzymes (e.g. α1-3/4 fucosyltransferases, α2-6 sialyltransferases). Glycosylation engineering of these hosts by stable transfection with genes encoding terminal human glycosyltransferases allows to obtain products with tailored (human tissue-specific) glycosylation in high yields.

Using site-directed mutagenesis, unglycosylated polypeptides can be successfully converted in N- and/or O-glycoproteins by transferring glycosylation domains (consisting of 7-17 amino acids) from donor glycoproteins to different loop regions of acceptor proteins.

The genetic engineering of glycoproteins and of host cell lines are considered to provide a versatile tool to obtain therapeutic glyco-products with novel/improved in-vivo properties, e.g. by introduction of specific tissue-targeting signals by a rational design of terminal glycosylation motifs.

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Authors and Affiliations

  1. Protein Glycosylation, GBF – Gesellschaft für Biotechnologische Forschung mbH, Mascheroder Weg 1, D-38124, Braunschweig, Germany

    Eckart Grabenhorst, Peter Schlenke, Susanne Pohl, Manfred Nimtz & Harald S. Conradt

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  1. Eckart Grabenhorst
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  2. Peter Schlenke
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  3. Susanne Pohl
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  4. Manfred Nimtz
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Correspondence to Harald S. Conradt.

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Grabenhorst, E., Schlenke, P., Pohl, S. et al. Genetic engineering of recombinant glycoproteins and glycosylation pathway in mammalian host cells. Glycoconj J 16, 81–97 (1999). https://doi.org/10.1023/A:1026466408042

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