Abstract: The present invention provides a novel method for the fast generation of high expression stable cell lines for the production of recombinant proteins with high efficacy of stable integration while using low selective pressure for only a short period of time. The method uses transiently expressed piggybac transposase to mediate stable integration of a transgene of interest flanked by the PB transposon termini.

Abstract: The present invention provides a novel method for culturing cells as well as a novel method for producing a recombinant protein by culturing cells at large scale (up to 1,500 L nominal volume and 750 L working volume), whereby an inflated bag provides a sterile, disposable cultivation chamber. The inflated bag is partially filled with liquid cultivation media and cells, and placed into a containment vessel. The containment vessel is positioned onto an orbitally shaken platform. The orbital shaking moves the containment vessel and thus the bag and induces thereby motion to the liquid contained therein (“shake mixing”). This motion (caused by orbital shaking) induces a dynamic force field that ensures cell suspension, bulk mixing, and oxygen transfer from the liquid surface to the respiring cells without damaging shear or foam generation.

Abstract: Culturing cells for the commercial production of proteins for diagnosis and therapy is a costly and time consuming process. The equipment required is expensive, and production cost are high. In order to provide commercially viable processes it is desirable to use cell lines which produce large quantities of product with each production run. However, most cells do not produce large quantities of desired product per se either because they do not produce a large quantity of product per unit of time (specific productivity) or because they do not survive long enough in the culture medium (time). Here, we identified that addition of a valproic acid compound to the culture medium increases overall (batch) yield and titer. More importantly, compared to the widely used sodium butyrate, batch yields using a valproic acid compound as a medium additive are significantly higher.