High cell density cultivation of mast cells in fluidized-bed and fixed-bed bioreactors

Noll T. High cell density cultivation of mast cells in fluidized-bed and fixed-bed bioreactors. Presented at the 17th European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies, Sweden

Bioprocess Development for the cultivation of human T-lymphocytes

Bohnenkamp H, Hilbert U, Noll T. Bioprocess Development for the cultivation of human T-lymphocytes. Presented at the 17th European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies, Sweden

Cultivation of human CMV-specific lymphocytes - an example for adoptive immunoterapy

Hilbert U, Biselli M, Noll T. Cultivation of human CMV-specific lymphocytes - an example for adoptive immunoterapy. Presented at the 17th European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies, Sweden

Metabolic flux analysis in mammalian cells - network modelling as an example for metabolic design in recombinant BHK cells

Paul W, de Graaf A, Marx A, Wagner R, Noll T. Metabolic flux analysis in mammalian cells - network modelling as an example for metabolic design in recombinant BHK cells. Presented at the 17th European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies, Sweden

Development of a generic transient transfection process at 100 L scale

We have developed a generic transient transfection process at 100 L scale, using HEK293-EBNA cells and PEI as the transfection reagent for the production of recombinant IgG. The process, including large-scale plasmid preparation, expression at bioreactor scale, capture, purification and, if necessary, endotoxin removal allows reproducible production of more than 0.5 g IgG for in vitro and in vivo studies. We compared the performance of two HEK cell lines, investigated the effect of conditioned medium, optimized the DNA:PEI ratio and implemented a feed strategy to prolong the culture time to increase product yield. The transient transfection protocol developed enables a closed process from seeding culture to protein capture. The challenge of performing a medium exchange before transfection at large scale is solved by applying a continuous centrifugation step between the seeding bioreactor and the production bioreactor. After 7–8 days the harvest and capture is performed in a one-step operation using a Streamline expanded bed chromatography system. Following a polishing step the purified antibody is transferred to the final formulation buffer. The method has shown to be reproducible at 10, 50, and 100 L scale expressing between 5 and 8 mg L−1 IgG