U. Scheffler

An integrated scale-down plant for optimal recombinant enzyme production by Pichia pastoris

An integrated bioprocess was created in a scale‐down production plant by developing a two‐stage enzyme production process with Pichia pastoris, containing a cell‐breeding reactor and a production reactor in combination with a three‐stage downstream process. To harvest the secreted enzymes, a disc separator and a cross‐flow microfiltration clear the broth from the cells. Purification with hydrophobic interaction chromatography removes other proteins, concentrates the product, and prepares the enzyme solution for lyophilization. Fully automated and broad observable multi‐stage parallel process courses have been developed using industrial process control systems and at‐line measurements for enzyme concentration and enzyme activity. Optimal process conditions were found by application of Design of Experiments (DoE) for the production process.

Advanced automation strategies for reliable, reproducible cultivation runs in a sequential/parallel operated multi-bioreactor plant

Abstract A designed multi-bioreactor plant, comprising a 5 l cell breeding BIOSTAT® Bplus system and a sixfold 1 l BIOSTAT® Qplus system, was equipped with different, interlinked automation structures for systematic process controlling and reliable optimization procedures. Benefits obtained were time saving, assurance and improvement of quality and cost reduction in bioprocess production. In this report the concept for moving beyond the state-of-the-art by creating a fully automated multi-bioreactor plant is explained. The efficiency of the approach is demonstrated by investigation relating the influence of the dissolved oxygen tension pO 2 in combination with the methanol concentration c S2M in Malaria vaccine production processes with Pichia pastoris.

Optimization of Recombinant Enzyme Production with Pichia pastoris in an Integrated Industrial Scale-down Production Plant

Abstract A scale down version of a multi component production plant, including a cell breeding reactor, a protein production reactor and a disc clarifier for enzyme separation is combined to an Integrated Bioprocess. Fully automated and global observable multistage parallel process courses are developed using industrial process control systems and atline measurements for enzyme concentration and enzyme activity. Optimal production conditions were found by application of DoE - Design of Experiment.

Monitoring and Control of Pharmaceutical Protein Production with Sequential Integrated Down Stream Processes

Abstract The methylotrophic yeast Pichia pastoris is used for expression of pharmaceutical proteins like 1-8del MCP-1 and vMIP-II. Here the proteins are secreted and these extra cellular products open new strategies in production processes. The combining of complex cultivations and primary down stream steps with cross flow filtration units leads to integrated bioprocessing. The development of optimised sequential production strategies in research and industrial scale needs on-line measurement of substrates and products up to the recombinant protein itself. Full automated processes are designed to analyse integrated filtration steps for modeling of cross flow units and combined bioreaction. The transfer of research work into industrial pilot plant scale is also demonstrated.

Computer Control Applications in Combined Waste Water Treatment Plants

Abstract During the past years one amplified research topic of the University of Applied Sciences in Hamburg was the purification of high nitrogen and methanol loaded waste water with simultaneous nitrification and denitrification processes. These include modern methods of bioprocess and automation technology as well as analytical techniques and simulation tools. The apparent processing was used to establish an advanced controlled waste water treatment process.

Integrated Bioprocess Development for Production of Recombinant Proteins in High Cell Density Cultivation with Pichia Pastoris

Abstract An integrated bioprocess is built up for production of an antagonist of recombinant monocyte chemoattractant protein-1 with Pichia pastoris. The process line based on full automated cultivation procedures includes micro-and ultra-filtration membrane modules for in-situ interactions as well as liquid chromatography methods for post cultivation opcrations.