Medizinische Universität Graz - Research portal
Selected Publication:
Grabenhorst, E; Schlenke, P; Pohl, S; Nimtz, M; Conradt, HS.
Genetic engineering of recombinant glycoproteins and the glycosylation pathway in mammalian host cells.
Glycoconj J. 1999; 16(2):81-97
Doi: 10.1023/A:1026466408042
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- Co-authors Med Uni Graz
- Schlenke Peter
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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 alpha2-3 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. alpha1-3/4 fucosyltransferases, alpha2-6 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.
- Find related publications in this database (using NLM MeSH Indexing)
- Animals -
- Carbohydrate Sequence -
- Cell Line -
- Glycoproteins - biosynthesis Glycoproteins - chemistry Glycoproteins - genetics
- Glycosylation -
- Glycosyltransferases - metabolism
- Humans -
- Mammals - metabolism
- Molecular Sequence Data -
- Mutagenesis, Site-Directed -
- Protein Engineering -
- Recombinant Proteins - biosynthesis Recombinant Proteins - chemistry Recombinant Proteins - genetics
- Find related publications in this database (Keywords)
- glycosylation engineering
- human fucosyltransferases
- human sialyltransferases
- N- and O-glycosylation
- host cell specificity
- recombinant glycoprotein expression