Bioprocessing is an essential part of many food, chemical, and pharmaceutical industries. Bioprocesses include the use of microorganisms and animal, plant mammalian cells for the production of new products. Recombinant proteins today represent an important part of pharmaceutical and biotechnological products. Whenever possible, they are expressed in microorganisms that have a high specific growth rate and high contribution, which is reflected in high productivity. The main task of the master's thesis is the development of recombinant protein production on a larger scale. The whole production process has to be characterized to enable efficient production process development. First we characterized bioprocesses in the shake flask. We determined the volumetric coefficient of oxygen transfer (kLa), the bacterial growth curve and the expression of recombinant proteins in the shake flask. We developed the processes for the isolation of the recombinant protein. With scale-up methods based on constant kLa and constant conditions of cultivation (medium, temperature, bioprocess type), we transferred the production of recombinant proteins to the stirred tank bioreactor. We did not reach the same growth kinetics and productivity at both scales. We achieved the similar trend of expression of recombinant protein and a similar cell productivity.
|
