Research

Energy and water consumption for buffer preparation can be substantial, especially if WFI is used. Whereas chromatographic process development can target minimal buffer use, the indirect ecologic impact of buffers in bioprocessing is completely unknown. This is because sustainability analysis and life cycle inventories are rudimentary and not fit-for-purpose to be used as development goal during process development. Traditionally, the reduction of buffer volumes is already part of the current implemented process development goals, as lower buffer volumes also correspond to lower costs and faster processes. Besides manufacturing of novel chromatographic formats (like filters, monoliths, etc.) this leaves very little room for further improvements in direct reduction of consumed buffers, but the sustainability influence of the buffer species and buffer strength is completely open for investigation, not addressed yet, and easy to implement in any scale as design and development goal. Buffer species, buffer preparation and buffer concentrations are currently selected by tradition, or by development goals completely unrelated to sustainability and we can tap into that underdeveloped area to select buffers and concentrations that are equally performant from a bioprocess viewpoint, but with a significantly lower ecological footprint. To tackle and quantify the influence of buffers and buffer preparation on downstream processing, we propose: (1) The generation of a comprehensive life cycle inventory of common buffer species used in biotechnological production (2) an assessment of the potential maximum savings through changes in buffer species and buffer concentrations that can be achieved by this approach (3) a fast to use tool for process development to assess the life cycle impact of buffers to implement a sustainability related key-performance-indicator that can be followed during process development (4) and finally the generation of physical demonstration use cases as a proof of concept.

Development of a process to produce recombinant Influenza Neuraminidase (rNA) antigen in the baculovirus system, and especially downstream processing/purification will be performed in collaboration between the Icahn School of Medicine at Mount Sinai and the University of Natural Resources and Life Sciences to optimize an affinity purification-based downstream process for production of his-tagged rNA which can be successfully implemented at the CMO Expression Systems to produce enough rNA for a Collaborative Influenza Vaccine Innovation Centers (CIVICs) phase I clinical trial. Furthermore, to develop a high-yielding tag-less purification process that would allow us to get sufficient protein yields for post-phase I clinical development and lastly, to perform testing of alternative rNA expression constructs and expression systems. Doing this work will enable us to test rNA vaccines in clinical trials and may also provide a commercial path forward for rNA protein-based vaccine development in general.

The vision of the CD laboratory is to move the production of rAAV for gene therapy from a cost-intensive, empirically-driven approach to a knowledge- and model-based process development and production. This requires a profound understanding of relationships such as target lines, therapeutic genes or process conditions interacting with one another and influencing the quality and quantity of rAAV. We will expand analyzes that allow accurate characterization of rAAV and contaminants. One of the most important tasks here will be the differentiation between therapeutic DNA-loaded and empty rAAVs, since different cell lines and different therapeutic genes lead to widely varying ratios between these variants. Product quality is currently being checked using post-process analysis. Such analyzes are time-consuming and costly, and require downtimes in the process. They only provide retrospective information and therefore cannot be used for process control. Therefore, sensors are being investigated that record important process parameters during the process and provide information about its course. This allows process monitoring and control, i.e. intervention in the process to ensure the desired quality. This approach not only increases the security of the processes, but also their efficiency. In order to develop a systematic understanding of important parameters and their interaction, different HEK target lines are examined and genome-wide analyzes of the cell response to virus production are carried out. Based on this, strategies to improve the rAAV yield and quality are developed. Scaling up optimized cell lines and processing strategies to production scale is a multi-step, time-consuming and costly process. The smallest scale currently available for the process development of rAAV production is the laboratory scale, which only allows a limited number of experiments due to the relatively high cost of materials. Only a miniaturized process development platform enables an integrated approach to investigate the relationships between process steps or upstream and downstream processing. This is therefore set up for the process development of rAAV and will include all relevant steps of cell cultivation and downstream processing. Finally, an optimized process is developed on this platform as an example and compared with a current process.