Toolbox approach for fast generation of stable CHO production cell lines from different hosts

Background Protein production in CHO cell lines is a highly dynamic process, where the yield limiting step can shift among transcription, translation, and secretion under different cell growth stages and culture conditions. In addition, the choice of CHO cell line has been shown to affect target protein quality and quantity. Further advancements in titer and specific productivity require elimination of cellular/process bottlenecks along the generation of stable production cell lines including vector design, clone selection, genetic engineering, and media and feed optimization. To overcome these limitations, we developed a toolbox for all process phases from transfection to production. This toolbox is based on a chemically defined media platform and includes pre-adapted CHO-K1 or CHO-DG44 cell lines, optimized vectors for single and multi-chain proteins as well as fine-tuned protocols. In addition, it enables the production of antibodies with specialized glycan structures (GlymaxX [1]) as well as clone engineering with regard to the expression and secretion machinery (CDC42).