Reduction of water and water-related energy consumption by in-situ media and buffer preparation on demand in continuous integrated

Figure 1 – (A) 3D printed single use device for feeding of dry powdered medium or buffer with its (B) top-, (C) front-, (D) and side view. A strong incentive for continuous integrated biomanufacturing is the improved economics compared to traditional batch wise biomanufacturing. The floor space and the size of unit operations are substantially smaller than in batch. This size reduction allows consequent implementation of single use technology, because single use equipment is not available at large scale biomanufacturing, e.g. reactor volumes above 2000 L or columns bigger than 100 L. However, a point often overlooked is that this transformation only “shrinks” the unit operations itself, while the necessary auxiliaries such as hold tanks, surge vessels and demand of process materials are unchanged or even drastically increased. Hence, the supply chain is facing an increase in demand of process materials and the necessity of handling significantly larger volumes. Media and buffer are always prepared in excess to mitigate the risk of failure for a campaign by running out of it. On a company level, but even more on a global level, this is an enormous waste of resources. We have previously shown that the water consumption is the major contribution to energy demand and CO2 emissions in a bioprocess. This link is described by the so-called metric WARIEN (WAter Related Impact of ENergy), which is also related to the metric PMI (Process Mass Intensity). The reduction and optimization of water consumption bears the highest potential to improve the environmental footprint of biomanufacturing, by reduction of energy consumption and CO2 emission. Here we show a single use device to continuously reconstitute chemically defined media on-demand and buffers directly from solids resulting in the same quality as by conventional batchwise preparation (Figure 1). The long-term operation over a duration of 12 hours demonstrated that such on-demand medium product is robust and precise. This technology with on-demand reconstitution directly from solids will make the repeated preparation of cell culture media and buffers and intermediate hold tanks obsolete which contributes significantly to the reduction of needed floor space. We present an economic and environmental analysis how the on-demand production improves economic and environmental footprint. This is exemplified by manufacturing of antibodies enzymes, hormones and growth factors. Preliminary economic analysis based on Biosolve and SuperPro models that were already built from data coming from the industry the amount of expenses that would be saved yearly on a global scale only by saving medium and buffer is almost 2.8 billion $ if we assume that 20% extra buffer is prepared for each of the bioprocess considered. This number is probably an overestimate but it\u27s just to demonstrate the potential and impact of this technology. The savings by reduction the floorspace are not take into consideration by this economic evaluation. The single use technology and process intensification towards integrated continuous biomanufacturing is a perfect marriage get improved sustainability of biomanufacturing

Elution profile from periodic counter current capture step as an on-line monitoring and control tool for perfusion bioreactors

Current trends in bioprocessing move towards the implementation of more on-line sensors such as Raman spectroscopy for titer monitoring in perfusion bioreactors. However, process performance data from one downstream unit operation can also be used to monitor and control the unit operation directly upstream. Despite several authors demonstrated a successful integration of continuous up-and downstream processes little attempts have been made to leverage the information derived from downstream processing as a real time feedback loop for upstream processing. We have developed a simple and robust approach in which protein A periodic counter current chromatography (PCCC) can function as an on-line monitoring tool for protein titer in continuous upstream fermentations. For a proof of concept, we exploit the fact that performance and binding capacities of state of the art protein A chromatography material do not significantly decrease throughout hundreds of cycles. Therefore, it is possible to predict the concentration of antibodies in the feed material from the elution pool and the volume loaded onto the column. We use the breakthrough curve during the interconnected phase of the PCCC, which is key for this approach. In the interconnected phase, the first column was loaded to 80% breakthrough, and the breakthrough curve modelled for a number of different concentrations in the feed material. Using the breakthrough curve, the time of the breakthrough can be modelled against the increase of product present in the feed stream, allowing the prediction of the concentration of antibody in the perfusion fermentation. This information feedback loop through the integration of PCCC and fermentation into effectively one single unit operation makes the titer determination in the fermentation obsolete, using the PCCC effectively as online monitoring tool itself. Future work after this proof of concept will include the prediction of protein A binding capacities through the lifetime of the resin and determination of accuracy and quantification limits the of interconnected units. Please click Additional Files below to see the full abstract

On demand, in-situ media and buffer preparation in continuous integrated biomanufacturing to reduce water consumption and CO2 footprint

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Solid polyethylene glycol precipitation: Potential cost reduction in antibody downstream processing

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