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Content archived on 2024-04-16

Control of recombinant protein glycosylation under defined cultivation conditions


Many products of major therapeutic value can be obtained only from animal cells grown in culture. The aim of this project is to improve the processes currently employed in this sector of biotechnology with special emphasis on the vectors and promoters used and on the quality of post translational modification.

The posttranslational modification of biotechnologically produced therapeutic/diagnostic glycoproteins will be studied and compared by applying different fermentation techniques for recombinant mammalian cell lines (BHK-21 cells). Cultivation systems used in animal cell culture technology will be evaluated including: perfused stirred tank and airlift reactor, immurement and entrapment techniques; influence of high density cultures, coated microcarriers; effect of energy sources as well as presence/absence of FCS/protein additives will be controlled.

Secreted products will be purified and the respective carbohydrate structures will be elucidated in detail. At the cellular level, studies will be carried out on the control of cellular nucleotide sugar level, formation of lipid intermediates as well as efficiency of initiation of N-and O-glycosylation. In addition, determination of the activity of cellular glycosyltransferases will allow a detailed description of the effect of different cell culture conditions on the cellular glycosylation machinery which determines the final carbohydrate structure of biotechnologically prepared glycoproteins.

Modelling of culture conditions will be performed leading to controlled posttranslational modification of products in long term culture. A procedure will be established enabling rapid batch analysis of purified glycoproteins with respect to their carbohydrate status ( "oligosaccharide-mapping" ) at the sub-milligram level.


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Gesellschaft für Biotechnologische Forschung mbH
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Mascheroder Weg 1
38124 Braunschweig

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