Scale down model in industrial cell culture processes – A powerful tool to ensure reliable production

Reliability and reproducibility of therapeutic protein production is one of the key objectives of GMP manufacturing with respect to product quality and supply to patients. But even in the age of chemical defined media, QbD and well characterized cell culture processes during manufacturing deviations can occur. While those deviations caused by human errors, equipment failures or process control (e.g. pH) can be assessed in a straightforward manner coming to satisfying results in an appropriate period of time, deviation caused by raw material lot variations are usually hard to detect. Development of risk mitigation strategies is complex since sophisticated analytical methods need to be applied and most often, additionally, vendors have to be involved in the root cause analysis. For risk mitigation change control assessments and QC testing covering a broad spectrum of parameters and quality attributes are applied before those materials are released for use in manufacturing. Even with this tight mesh of control quite a number of incidences can be reported, demonstrating that deviations based on raw material issues have a huge impact on our business. Here we present several case studies, where neither QC testing nor vendor and customer change control could predict the deviating process behavior observed when these materials had been used in the cell culture process at production scale. As a consequence cell culture use tests by validated scale down models have been introduced for several raw materials in the meantime. These test procedures will be presented

Elimination of Fetal Calf Serum (FCS) in industrial cell culture media for manufacturing of diagnostic Anti-bodies

Here we demonstrate the suitability of different substitutes for the use of fetal calf serum (FCS) in cell culture media. Due to decreasing availability of FCS on the global market whilst growing demand of cell culture media to support Anti-body production for more than 200 different invitro-diagnostic kits media development efforts had to be taken. A benchmark of different substitutes, such as NBS (new born serum), human thrombocytes lysate, and chemically defined media (CDM) will be presented. Case studies from lab scale (10Ltr) up to full production scale (2,000Ltr) will be shown

Improving the batch-to-batch reproducibility in microbial cultures during recombinant protein production by guiding the process along a predefined total biomass profile

In industry Escherichia coli is the preferred host system for the heterologous biosynthesis of therapeutic proteins that do not need posttranslational modifications. In this report, the development of a robust high-cell-density fed-batch procedure for the efficient production of a therapeutic hormone is described. The strategy is to guide the process along a predefined profile of the total biomass that was derived from a given specific growth rate profile. This profile might have been built upon experience or derived from numerical process optimization. A surprisingly simple adaptive procedure correcting for deviations from the desired path was developed. In this way the batch-to-batch reproducibility can be drastically improved as compared to the process control strategies typically applied in industry. This applies not only to the biomass but, as the results clearly show, to the product titer also