Boehringer Ingelheim International Gmbh

Jochen Schaub

Markus Mueller

Torsten Schulz

Christian Bernloehr

Jennifer Koenitzer

Peter Schorn

Christoph Clemens

Marie Link

Torsten Wilhelm Schulz

The present invention relates to the identification of a genomic integration site for heterologous polynucleotides in Chinese Hamster Ovary (CHO) cells resulting in high RNA and/or protein production. More specifically it relates to CHO cells comprising at least one heterologous polynucleotide stably integrated into the S100A gene cluster of the CHO genome and to methods for the production of said CHO cells. Further, the invention relates to a method for the production of a protein of interest using said CHO cell and to the use of said CHO cell for producing a protein of interest at high yield. Integration within these specific target regions leads to reliable, stable and high yielding production of an RNA and/or protein of interest, encoded by the heterologous polynucleotide.

Integration sites in CHO cells

The present invention provides a basal cell culture medium and a feed medium with novel amino acid ratios and/or iron choline citrate as iron carrier that result in improved performance of mammalian cell culture processes, such as CHO cultivation and protein production processes, in particular in increased product titer (e.g. of monoclonal antibodies). Also provided are methods for culturing mammalian cells and producing a protein of interest using said basal cell culture medium and optionally feed medium. The invention also provides for a medium platform that comprises (i) the basal cell culture medium and (ii) the feed medium.

Cell culture medium

Biopharmaceutical process development with recombinant protein producing mammalian cells has realized a tremendous increase in both productivity and product yields in the past years. These achievements can be mainly attributed to the advancements in cell line development, media, and process optimization. Only recently, genome-scale technologies enable a system-level analysis to elucidate the complex biomolecular basis of protein production in mammalian cells promising an increased process understanding and the deduction of knowledge-based approaches for further process optimization. The present invention describes a method for a rational cell culturing process using such a knowledge-based approach.

Method for a rational cell culturing process

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Biberach an der Riss, Germany

Jochen Schaub