Using whole process models to go beyond Cost of Goods in terms of productivity (based on facility volume), material intensity, and total energy efficiency. By focusing on these metrics the user can rapidly assess the impact of a new process, technology compared to the status quo. Providing a guide to process/facility options that minimize environmental impact and maximize the ROI whether looking at a new operation or retrofit. By way of example, comparisons are made between standard fed-batch processes and intensified process options that include perfusion and continuous downstream operations at different scales.

We have developed a steady-state perfusion process of high cell density at 100 x 1E6 cells/mL in 200 mL scale bioreactor integrated with a 3-steps continuous purification. This process was successfully transferred to pilot scale (30 L bioreactor ) and generated 30 g/L purified antibody per day. The presentation will focus on the integration and the upstream process development. 

As current biological product pipelines become more diverse, product demand and cost pressures are increasing, manufacturers are shifting towards process intensification. By altering unit operations, processes, or even facility type, industry can identify areas of potential improvement to increase productivity, reduce timelines, downsize process footprint, lower cost of goods, and/or unlock additional manufacturing flexibility. The presentation will focus on these options whilst considering a more “sustainable” future.

For complex proteins, fed-batch cultivation might lead to product degradation, while shifting to perfusion as production process yields high-quality product. Our case study shows the successful transfer of an optimized perfusion process for a difficult to express protein expressed in fully human host GEX to a fed-batch process with an n-1 perfusion step to increase initial cell density. Product quality was still satisfying while yield exceeded our expectations.

Process intensification (PI) of mammalian upstream processes is sought to encounter upcoming demands of biopharmaceuticals due to optimized productivities and reduced plant’s footprints. Rational approaches are required to achieve this. This talk demonstrates a strategy to implement an intensified based on a conventional CHO cell fed-batch (FB) platform process (< 2000 L) using N-1 perfusion for subsequent high inoculation FBs. Additionally, butyric acid was used to increase cellular productivities. Ensured by maintained CQAs and detailed process insights from metabolomic and trancsriptomic responses towards PI the space-time yield of the IgG was ultimately and successfully doubled.

The discussion will be focused around the exploration of multiple techniques and their possibilities to secure tight control of the major critical quality attributes (CQA) and different PAT tools for measuring different attributes in (near) real-time. Feasibility study of PAT tools covered in this talk demonstrates the technology for in-line measurements in fed-batch process steps and potential of their use in a continuous process.

This presentation covers 3 consecutive projects, following CPI's progression in establishing and developing continuous processing capability with integrated PAT-based controls. This talk will follow the progression of a mAb platform process, from establishing a downstream only continuous process with basic automation, to the ongoing project that will utilise real-time model-based advanced process control and will include integrated upstream perfusion.

Genome-integrated, as well as growth-decoupled E. coli expression systems, enable continuous protein production. Efficient implementation requires suitable process strategies for cultivation and product recovery and purification. The presentation will show two case studies inclusive of an economic evaluation with standard fed-batch as benchmark.

Downstream processing of mAb fragments typically starts with capturing by protein L affinity chromatography. 

In this talk performance parameters of several Protein L chromatography media will be compared and their influence on process economics will be evaluated.

Further, we will demonstrate how transferring from a conventional batch mode to continuous affects the productivity of the antibody fragment capturing process.

In conventional perfusion processes, feedback control algorithms are applied to achieve constant process performance and cell density by tightly controlling feed, harvest, and bleed flow rates. Such controllers lead to increased deviations from process set points over time. A single prediction controller (SPC) enables the compensation of process variations that normally would be transferred to adjacent units in integrated continuous bioprocesses (ICB). Steady-state operation over long periods eases the integration of the bioreactor with subsequent capture units. This is of particular interest when the capture step is sensitive to fluctuations in product concentration and product streams.

Continuous manufacturing and platform processes are the “hot” topics of bioprocessing both in research and industry. As the upstream moved forward, downstream purification processes are today the bottleneck to overcome. Precipitation is a simple unit operation and by far easier to implement when compared to chromatographic processes. A proof of concept is presented: a PEG capture step with CaCl2 pre-treatment of cell culture supernatant, combined with a ZnCl2 washing step.

Ultra- and Diafiltration outcomes are strongly influenced by product and matrix characteristics. Filtration may take longer than expected and the target formulation pH is difficult to obtain. We present a hybrid model structure consisting of ML-supported flux predictions and mechanistic models for buffer and pH prediction, giving an in-depth understanding of this crucial process unit. We present a holistic filtration model that uses process simulations to meet the desired CQAs. The hybrid model is implemented in our modeling toolbox, including appealing visualization, simple import of parameters by drag-and-drop, and easy-to-use slider bars to see the effect of parameter variations instantly.

Viral safety is a Critical Quality Attribute for mAbs purified by Protein A chromatography.
Murine Minute Virus (MMV)-spiking experiments were performed at small scale on BioSC™ Lab in order to compare multicolumn (SMCC process) vs batch-column chromatography.
It was confirmed that viral clearance was similar in both cases, securing the use of multi-columns for downstream processing.

The presentation will also include an overview of the largest setting of the BioSC™ platform.

There is significant need to improve existing AAV manufacturing platforms to achieve robust, high-yielding, scalable, and cost-efficient processes. At Ultragenyx we employ scalable AAV production processes using mammalian producer cell lines that eliminate the high cost of goods associated with the traditional AAV platforms. In the following presentation, we demonstrate how we have improved on our existing platform process through process intensification to achieve a several-fold increase in overall volumetric AAV yield. Through process intensification, we have increased our AAV platforms productivity to =3E11 GC/mL (=3 E14 GC/L).

With the first products now to market, mRNA represents a disruptive technology with broad applications and the potential for a platform manufacturing approach. This talk will outline our experiences evaluating and developing a modular manufacturing platform for mRNA, and view to how such processes could be intensified.