Applikon Biotechnology’s leading role in continuous processing

Applikon Biotechnology’s leading role in continuous processing

applikon biotechnology 150x48

Products & Services | Applikon Biotechnology B.V.


Applikon Biotechnology is playing a leading role in the emerging move across the biopharma industry to embrace continuous process (CP) bioprocessing methodologies.

CP, already conventional in other fields of manufacturing, appears to be an idea those time has come for pharma, due to its compelling advantages that include enhanced product quality, greater efficiencies and more rapid scalability.

Applikon is providing solutions for continuous processing, including compact bioreactors, perfusion and cell separation systems and the advanced and highly scalable control systems needed to automate and closely monitor continuous processes.

The Trend to Continuous Processing

While most biopharmaceutical production continues to be based on batch-fed processes, continuous processing (CP), largely via perfusion, is rapidly gaining traction.

Amgen, Genzyme and many other leading biotech producers are now pursuing technologies that streamline biopharmaceutical manufacturing via system-wide use of CP.

In biopharmaceutical manufacturing, CP is based on bioreactors running on continuous cycle as opposed to “batch” production, where a specific quantity of drug is produced in a tank during a single discrete manufacturing cycle.

Where production scale batch processing tends to feature large bioreactor tanks (10,000 liters or more), CP emphasizes smaller bioreactors continually feeding media to specialized chromatography processes to separate the product from the surrounding liquid.

Continuous Processing Advantages

The major advantages of CP are lower equipment costs and smaller quantities of cell media, leading to a reduced overall footprint, eliminating redundant holding containers and clarification systems. CP also delivers a series of other benefits:

Improved Product Quality
Product quality is enhanced since media nutrients are constantly maintained and cells don’t experience the drop-off or lags, often encountered at end of a batch run. Further, processing delays are greatly reduced as product constantly flows from one step to the next, eliminating major causes of product degradation and contamination.

Enhanced Scalability
Scalability is becoming a key industry issue as drug shortages become of higher concern to the FDA and EMA, etc. Since CP production is primarily governed by durations rather than volumes, production can simply be ramped up by extending campaigns and adding further compact CP units. CP also supports easier technology transfer, since the systems used in laboratory R&D tend to the same as those employed on production floor.

Increased Profitability
Continuous processing enhances profitability in two ways: by allowing more efficient and economical facility design and utilization and by reducing the cost of goods through reduced footprint, lower media and energy consumption, enabling increased automation with consequent reduction in service requirements and operator time.

Perfusion in Continuous Processing

While few approved biopharmaceutical manufacturing processes currently use CP that does not mean a switchover is particularly problematic. Many existing and GMP-compliant biopharmaceutical manufacturing processes are inherently compatible with fully continuous processing methodology.

One reason for the easy fit is the current use of perfusion bioreactors. In perfusion mode, the culture is maintained over much longer periods by continuously feeding the cells with fresh media and removing a fraction of the spent media and soluble product. Perfusion cell culture is well tested within the biopharma industry having first been introduced in the early 1980’s as a means for improving the productivity of cell-culture processes and safely capturing unstable/labile proteins that degrade under batch and fed-batch process conditions.

Now increasing interest in CP for biotech is shifting attention to perfusion’s inherent advantages for upstream cell-culture processes. This transition is being unlocked with further developments in automation and downstream of the perfusion bioreactor system, where protein can in theory be continuously separated from the steady stream of harvested liquid media.

The major challenges in adopting CP concern new forms of in-process testing and release approaches, along with the need to adapt more robust and adaptive closed loop control systems.

CCTC for Continuous Downstream Processing

New solutions being developed in the area of perfusion include Chromatan Corporation’s patented Continuous Countercurrent Tangential Chromatography (CCTC) technology that overcomes many of the limitations of batch columns without the drawbacks of multi-column systems.

The CCTC platform is a true moving bed technology that runs at steady-state without utilizing columns. The novel design of the system enables short residence times, consistent product concentration and quality, as well as easier implementation of advanced Process Analytical Technology (PAT) and process control.

The patented CCTC system includes all of the traditional chromatography steps such as bind, wash, elution, strip, and regeneration, but these operations are conducted on a moving slurry instead of a packed column. Each step is made of multiple stages arranged in a countercurrent configuration, each of which is composed of a static mixer and a hollow fiber membrane module. In a CCTC process, the resin slurry flows through these modular stages, while all of the chromatographic operations are performed simultaneously and at steady state. (Figure 1).

The CCTC system has been shown to operate continuously with 5-10x greater productivity vs. batch columns, enabling a significant reduction of resin volume compared to batch column requirements.

CCTC Perfusion Case Study

In Phase I of an NIH grant, Chromatan teamed up with Applikon to produce a commercial mAb. Experiments were designed to demonstrate effective use of a perfusion bioreactor run for long-term CCTC hydrodynamic testing and CCTC purification testing.

An Applikon 3L bioreactor with ez-Control and BioSep perfusion device were used to perform the bioreactor run.

High cell density (50-60 x106 cells/mL) was obtained and maintained for ~ 30 days (Figure 2). The test results obtained with the BioSep demonstrate the suitability of this technology for long-term continuous perfusion with the added benefit of improving the quality of the cell culture by preferentially removing dead cells.

Applikon Continuous Processing Solutions

Applikon Biotechnology offers a range of solutions that support advanced CP applications, including advanced DeltaV based control systems, advanced bioreactors and cell separation systems. These include:

The Applikon BioSep system provides a uniquely effective solution for cell retention in high-density perfusion processes.

Based on acoustic resonance technology, BioSep offers all the benefits of traditional devices but without their inherent fouling and clogging problems and other limitations by using ultrasonic sound waves to separate cells, rather than a physical mesh or membrane. The BioSep is directly mounted onto the bioreactor head plate (brand independent) and can be set up to allow for continuous operation, semi-continuous operation or alternative strategies of cell recirculation.

Proven under cGMP conditions and available for harvest rates from 100ml/day (mini BioSep) up to 1000L/day, the system is easy to install and to operate and can be used in combination with any brand of bioreactor.

Applikon has long been noted for the quality of its laboratory bioreactors, with the bench-scale MiniBio2 range as the latest iterations offering ultra-compact continuous processing at reactor volumes between 250 ml and one litre.

In MiniBio 2, autoclaving, installation and interconnection are all simplified resulting in noticeable time savings and efficiency gains in setting up these systems.

The MiniBio2 is a true scale down of the classic laboratory scale bioreactor with the same versatility and flexibility that allow it to be tailored to any research process, while reducing media costs and maximizing premium bench space.

Integrated Software solutions
Applikon controllers run powerful on board software, but the CP market also demands automation and integration of PAT. Applikon provides two unique solutions to meet this need.

Lucullus Process Information Management System (PIMS) integrates functionalities beyond the classic SCADA solutions. Creating and planning of recipes, reactor hardware allocations, DOE, media preparation, data analysis, automatic reporting and modeling are all built-in to one comprehensive software platform. Bringing all the data and functions in to one software solution saves time and reduces risk, critical capabilities for a system running in CP.

V-Control from Applikon is based on Emerson’s industry-leading DeltaV process automation platform but offers both bench scale or manufacturing scale implementation. Allowing the user accurately to control and reduce variability in key process parameters, using the familiar DeltaV operator user interface to automate an entire continuous process.

Soroosh Soltani1, Amit K. Dutta1, Dmitriy Fedorenko1, Jasmine Tan1, John Chorazyczewski2, Ann D’Ambruoso2, Andrew L. Zydney3, Oleg Shinkazh1,*

1 Chromatan Corporation, 200 Innovation Blvd., Suite 260B, State College, PA 16803
2 Applikon Biotechnology, 1180 Chess Dr, Foster City, CA 94404
3 Department of Chemical Engineering, The Pennsylvania State University, University Park, PA 16802


Click on Applikon in Continuous Processing for more information.
Click on Applikon Biotechnology B.V. to contact the company directly.

Figure 1: Schematic of Chromatan CCTC chromatographic perfusion continuous process

Figure 1: Schematic of Chromatan CCTC chromatographic perfusion continuous process

Figure 2: Cell density and viability data from Chromatan/Applikon NIH grant study: High cell density (50-60 x106 cells/mL) obtained over ~ 30 days

Figure 2: Cell density and viability data from Chromatan/Applikon NIH grant study: High cell density (50-60 x106 cells/mL) obtained over ~ 30 days

Applikon BioSep acoustic resonance cell retention device for high-density perfusion

Applikon BioSep acoustic resonance cell retention device for high-density perfusion

Applikon MiniBio2 benchtop scale laboratory bioreactor

Applikon MiniBio2 benchtop scale laboratory bioreactor

Supplier Information
Supplier: Applikon Biotechnology B.V.
Address: Heertjeslaan 2, 2629 JG Delft, The Netherlands
Tel: +31 10 208 3555
Fax: +31 10 208 3505

Comments are closed.