Boehringer Ingelheim Pharma Gmbh & Co Kg

Boehringer Ingelheim International Gmbh

Eric Becker

Lore Florin

Hitto Kaufmann

Monilola Olayioye

Angelika Hausser

Joey M Studts

Tim Fugmann

The invention concerns the field of cell culture technology. The invention describes production host cell lines comprising vector constructs comprising a CERT S132 A expression cassette. Those cell lines have improved growth characteristics and high CERT S132A expression levels. The invention especially concerns two cell lines deposited with the DSMZ under the number DSM ACC2989 (CHO/CERT 2.20) and DSM AC-C2990 (CHO/CERT 2.41). The invention further concerns a method of generating such preferred production host cells and a method of producing proteins using the two cell lines deposited with the DSMZ under the number DSM ACC2989 (CHO/CERT 2.20) and DSM ACC2990 (CHO/CERT 2.41).

CHO/CERT cell lines

The invention concerns the field of cell culture technology. The invention describes production host cell lines comprising vector constructs comprising a DHFR expression cassette. Those cell lines have improved growth characteristics in comparison to DHFR-deficient or DHFR-reduced cell lines such as CHO DG44 and CHO DUKX-B11. The invention especially concerns two cell lines, a representative of each cell line is deposited with the DSMZ under the number DSM ACC2909 (CHOpper&#xae; Discovery) and DSM ACC2910 (CHOpper&#xae; Standard). The invention further concerns a method of producing proteins using the cells generated by the described method.

Production host cell lines

The invention concerns the field of cell culture technology. It concerns a method of improving cell growth, especially the growth of biopharmaceutical producer host cells. The invention further concerns a method of producing proteins using the cells generated by the described method.

Cell growth

The invention concerns the field of protein production and cell culture technology. CERT is identified as a novel in vivo PKD substrate. Phosphorylation on serine 132 by PKD decreases the affinity of CERT towards its lipid target phosphatidylinositol 4-phosphate at Golgi membranes and reduces ceramide transfer activity, identifying PKD as a regulator of lipid homeostasis. The present invention shows that CERT in turn is critical for PKD activation and PKD dependent protein cargo transport to the plasma membrane. The interdependence of PKD and CERT is thus a key to the maintenance of Golgi membrane integrity and secretory transport.

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Hochdorf, Germany

Eric Becker