There has been an increasing trend in developing unit operations that can been utilised in a continuous fashion, although fully continuous end to end operations are not fully realised yet there are opportunities to use these unit operations in a hybrid mode to increase output, reduce footprint and reduce cost of goods. Data from experiments to show effectiveness of these approaches.

An intensified perfusion process for production of a therapeutic monoclonal antibody was developed and scaled to 100 L clinical and 500 L commercial scales. The baseline process was developed in 10 L bench top bioreactors with stainless steel alternating tangential flow (ATF) cell retention systems. The process consisted of a 12 day growth phase followed by a 48 day harvest phase. Cell densities of >120 Mvc/mL were sustained with high culture viability. Productivities of >3 g/L were maintained throughout the harvest phase. The process was successfully scaled up to a 100 L single use bioreactor with dual ATF6 filters for clinical manufacturing. To verify that the process would perform similarly at commercial manufacturing scale, a proof of concept run was conducted in a 500 L single use bioreactor with dual ATF10 filters. Biomass concentration culture viability, and productivity were comparable across scales. A full 60 day campaign in a 500 L bioreactor would generate over 70 kg of product in the clarified harvest. These studies demonstrate that intensified perfusion processes developed in benchtop bioreactors can be successfully reproduced at scales relevant for manufacturing.

PAK BioSolutions newly released system performs end-to-end continuous purification of biopharmaceuticals in a fully-automated manner and is highly configurable to match any purification process design.  The systems capabilities were demonstrated with continuous purification of a 50L perfusion bioreactor over 14 days.  We will present product quality and bioburden data from capture chromatography, virus inactivation, filtration, and polishing membrane operations performed in series simultaneously with a single piece of equipment.  

Complexity grows when we move from batch to continuous processing and a unified platform ensures you have the right tools for real-time processing, optimizing processes, and predicting yields to produce the best quality product. Learn how to:

• Generate insights by bringing in data real-time from disparate sources

• Effectively adopt and integrate AI in a qualified manner

• Find patterns that may lead to deviations and take preventative action

• Future-proof for what’s ahead

Continuous integrated manufacturing can be achieved of biopharmaceuticals has been achieved by converting batch unit operations into a pseudo continuous operation. Here we demonstrate a real, fully continuous operation and a monitoring and control concept for production of antibodies. The perfusion culture is directly linked to a continuous precipitation and virus inactivation in a single line. The economic advantages for such a process will be also discussed.

Continuous Biomanufacturing is the highest level of integrated bioprocessing available to deliver drug products to patients.  These qualities enable unique functionality and platforms capable of rapid deployment delivering agile and accelerated timelines from development to on-demand manufacturing.  The drivers for adoption of CBM relate to improved productivity, reduced plant footprint, and favorable lifecycle considerations.  These come at the cost of more sophisticated supervisory controls, to enable consistent quality and in-process continued verification.  Companies considering CBM platforms must be ready to support their applications with robust control strategies that are very sound in evidence-based considerations as well as risk-based justifications.

This talk will focus on the upstream assessing the impact of strategies for process intensification:​

• Concentrated N-1 seed, perfusion, what are the optimum configurations
• High turndown bioreactors in the seed train
• Impact of concentrated Fed batch
• The implications of deploying large volume single-use bioreactors, upto 6000L

The outcomes of modelling will provide insights into the benefits and the issues that need to be addressed for a successful financial outcome.​

Manufacturing of biopharmaceuticals requires high quality water. We combined economic and ecological modeling to assess energy consumption of water production from tap to waste and defined the WAter Related Impact of ENergy (WARIEN) metric to directly correlate the amount of CO2 emitted per kg biopharmaceutical. The water related costs per kg product and the WARIEN correlate and therefore can be used as a design criterion for biopharmaceutical process development.

Mathematical model of process can help for process simulation, prediction, monitoring and/or control. We have developed modelling methodologies suitable for perfusion or fed-batch processes, including amino acid metabolism or glycosylation. We will illustrate the prediction power of these models in high cell density perfusion runs. The models are based on the measurements of the extracellular metabolites, and include an objective and automatic determination of the used pathway and the kinetics.

A perfusion process is particularly well suited to the biomanufacturing of lentiviral vectors due to their labile nature. A comparison of the titers obtained in different modes of operation will be summarized. We favor the use of stable producer cell lines over transfection, and we demonstrate that generating the most stable cell line possible is worthwhile. Finally, our efforts towards a semi-continuous process from upstream to downstream will be presented. 

For over 10 years, PendoTECH Single Use Sensors have been used extensively both in the laboratory for process development and in GMP processes to enable elimination of cleaning and facilitate rapid turnover of process equipment. Their primary applications are for measurement of pressure, temperature, conductivity, UV, and turbidity in a range of upstream and downstream applications. PendoTECH is offering a platform to enable all of its sensors to smoothly transition to use for Bioprocessing 4.0.

We demonstrate a new micro/nanofluidic system for continuous and automatic monitoring of protein product size and quantity directly from the culture supernatant during a high-cell-concentration CHO cell perfusion culture. The continuous-flow and fully automated operation of this nanofluidic protein analytics reduces design complexity and offers more detailed information on protein products than offline and batch-mode conventional analytics. Moreover, chemical and mechanical robustness of the nanofluidic device enables continuous monitoring for several days to a week. This continuous and online protein quality monitoring could be deployed at different steps and scales of biomanufacturing to improve product quality and manufacturing efficiency.

Real time quality attribute monitoring in bioprocess through process analytical technology (PAT) is a vital necessity for precise process control, and better process understanding. On-line Multi-attribute Chromatography (MAC) is one of the PAT tools that allows measurement of few critical quality attributes in real time. Here we describe an On-line MAC platform featuring Two-Dimensional Liquid Chromatography (2DLC) and automated-sampling device for real time monitoring of charge variants and aggregation in production bioreactor. The workflow involves automated-extraction of a cell-free sample from the bioreactor followed by Protein A purification in the first dimension and subsequent analysis for quality attributes such as charge variants and aggregation in the second dimension.This technique can be applied for monitoring from multiple bioreactors with different protein modalities. Additional capability is the potential as a tool for further analysis of charge variants via mass spectrometry.

Digital twins will play crucial role in continous biomanufacturing, because product quality needs to ensure in a time continuum. The digital twins need to be set up and also maintained in an efficient but also robust manner, so that they keep up with the evolution of the process life cycle. This contirbution will show new workflows how to generate and adapt digital twins for upstream and downstream applications.

Pall’s mPath IoR concentration monitor provides accurate inline continuous measurement of product concentration in bioprocess fluids, enabling more rapid development and characterization of biotech processes. These measurements enable scientists to quickly see the impact of process changes on product yield, allowing fast determination of optimal process parameters

mAb purification must ensure the removal of host cell proteins (HCPs), endotoxins, mAb fragments and aggregates. Commercial adsorbents can struggle to remove particular “high-risk” HCP impurities that pose direct threat to health. We demonstrated a novel adsorbent – LigaGuard^TM – that captures HCPs in flow-through mode and enables continuous mAb purification from CHO, HEK, and Pichia fluids. We demonstrated >92% recovery, >95% purity, HCP-LRV ~ 2, and clearance of “high-risk” HCPs.

Microbes in biofilm forms usually express magnificent potentials for boosted product secretion through profound genetic alterations during the immobilization process. Biofilm reactors provide the controlled environments for filament and biofilm formers to express these great advantages without the associated transport phenomena shortcomings and scale-up issues. The audience will take on the fundamentals of biofilm reactors for production of value-added products and see the latest most cutting-edge technologies based on them. Moreover, they will be presented the perspective how biofilm reactors may be find their way into manufacturing suites of recombinant biopharmaceuticals.

Perfusion cell culture has emerged as a promising technology for continuous biologics’ production. Nevertheless, the necessity of cell bleed or growth inhibitors in traditional steady-state perfusion results in unnecessary product loss. Consequently, we have developed an osmolality-based dynamic perfusion process utilizing compartmentalized media concentrates to support ultra-high cell density (>100e6 cells/mL). This process eliminates the need for cell bleeds since it can achieve substantial productivity improvement in the length of conventional mammalian upstream process. Pilot-scale application further demonstrates a 10-fold enhancement in productivity compared with fed-batch operations, revealing its potential as an innovative approach for next-generation continuous manufacturing.

The 2 mL Erbi Breez micro-bioreactor system (developed by Erbi Biosystems, Woburn, MA) utilizes microfluidic technology to integrate the reactor chamber, pump mechanism, process sensors, and perfusion unit operations into one single-use cassette. The small footprint, system flexibility, and integrated controls make it a promising candidate for an intensified perfusion scale-down model. Our primary evaluation generated results that are comparable to the bench scale perfusion process.