Datensatz

Zusammenfassung

To better understand bioprocesses for targeted optimisation of productivity proteome analysis throughout the production process can be a helpful tool. Accumulated data of such an investigation could be used to identify for example intracellular effects of different media or bioreactor conditions in a bioprocess. Our goal is the optimization of a mammalian cell-based influenza virus vaccine production process, which typically uses transformed cell lines such as MDCK, Vero or PER.C6 cells. However, as most protein databases contain mainly human proteins we look for an appropriate human cell line for a possible comparative proteomic investigation of virus-host cell interactions. In this work we have screened the metabolism and virus productivity of three different human cell lines: a gastric carcinoma cell line (NCI-N87), a hepatocellular carcinoma cell line (HepG2) and a lung carcinoma cell line (A549), to identify the best candidate for our proteomic approach. Therefore, experiments on cell metabolism during cell growth and virus infection in different media and cultivation vessels were carried out. Here, we present first results, comprising the determination of the metabolite concentrations of glucose, lactate, glutamine, glutamate and ammonia in the different media to follow the energy metabolism as well as to identify media limitations and growth inhibition effects. Additionally, virus titers in different media and different multiplicity of infection (m.o.i.) were compared for infection with human influenza A. On the basis of these data, A549 was selected for future studies applying 2DDIGE technology to compare protein composition of the selected human cell line with the proteome of MDCK cells during an influenza vaccine production process.