Single-use bioreactor for process intensification
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1 USD2941 (1) icellis Bioreactor Single-use bioreactor for process intensification Highlights and Benefits Process simplification, from pre-culture to final product The first fully-integrated, single-use bioreactor (integrated perfusion with disposable, pre-installed calibrated probes and no additional oxygen device needed) Straightforward implementation for faster process development Linear scalability from R&D to manufacturing Compact high-cell-density, fixed-bed bioreactor providing a significant increase in volumetric productivity vs. traditional stirred tanks Dramatic decrease in operational costs and capital investments
2 Overview The icellis bioreactor is the world s first fully-integrated, high-cell density bioreactor designed to simplify your process by combining the advantages of single-use technologies with the benefits of a fixed-bed system. The compact system designed for quick implementation and ease-of-use represents a new generation of single-use bioreactors. Central to the icellis bioreactor technology is the use of a compact fixed-bed, filled with custom macrocarriers. This matrix is made of medical grade polyester microfibers and provides in only 25 L volume up to 500 m 2 available area for cell growth. This is the surface-equivalent of 3,000 Roller Bottles (at 1700 cm 2 each). The icellis bioreactor is provided with pre-packed macrocarriers. Compared to a standard stirred-tank bioreactor using microcarriers, the icellis technology leads to a dramatic simplification of the cell culture process. It alleviates delicate and time-consuming manual operations, reducing the overall process time and making the cell culture process more robust. With biomass multiplication occurring in the fixed-bed, icellis bioreactors can be inoculated at very low density. The number and volume of pre-culture steps are reduced considerably; 20 Roller Bottles (1700 cm 2 ) are sufficient to inoculate an icellis 500 bioreactor (with 500 m 2 of available surface to the cells). As a result of this process simplification, manual operations and associated costs are greatly reduced. Evenly-distributed media circulation is achieved by a built-in magnetic drive impeller, ensuring low shear stress and high cell viability. The cell culture medium flows through the fixed-bed from the bottom to the top. At the top, the medium falls as a thin film down the outer wall where it takes up O 2 to maintain high KLa the in the bioreactor. This unique waterfall oxygenation, together with a gentle agitation and biomass immobilization, enables the compact icellis system to achieve and maintain high cell densities equaling the productivity of much larger stirred-tank units. Process simplification, from vial thawing to final product, is at the heart of the icellis platform. As cells are immobilized in the fixed-bed, and the icellis system operates in perfusion mode, no centrifugation is needed to harvest the cells. This benefit further simplifies the downstream process. Media flow and average media in the icellis 500 bioreactor (60-70 L) Features and Configurations Microscopy view of MDCK cells immobilized on icellis system carriers Integrated mixing system for evenly-distributed media circulation and low shear stress Specialized carriers specifically adapted to adherent cell cultures Unique waterfall media oxygenation for high oxygen transfer Single-use bioreactor made from USP Class VI rigid plastic to ensure process reliability Modular height of fixed-bed from 2 cm to 10 cm offering several configurations of small and large scale The icellis bioreactor is available in two formats: The icellis nano system for feasibility studies and small-scale production The icellis 500 system for industrial scale manufacturing (up to 500 m 2 ) The icellis 500 bioreactor (500 m 2 ) is equivalent to 3,000 Roller Bottles (1700 cm 2 each) The icellis 500 bioreactor on its fully-integrated skid 2
3 Table 1 Configurations of icellis bioreactors at small and manufacturing scale Bioreactor Fixed-Bed Fixed Bed Bioreactor Surface Area (m 2 ) Bioreactor Diameter (mm) Height (mm) Volume (L) Volume (L) Low Compaction High Compaction icellis nano icellis nano icellis nano icellis 500/ icellis 500/ icellis 500/ icellis 500 Bioreactor Components 1 Probe supports (T, ph, D.O.) 2 Outer shell, top section with 8 ports (5 IN/3 OUT); Media IN/OUT, Feed IN/OUT, NaOH, Gas IN/OUT, Cell/Virus 3-5 Fixed-bed support integrating pump housing (5) and filled with macrocarriers (4). Top grid to retain carriers (3) 6-7 Integrated magnetic-driven impeller (6) and pump housing ring (7) 8-9 Outer shell, bottom section including double-jacket (9) Process Control Systems Startup Requirements Power Requirement 500 W 3500 W (2) Power Supply VAC VAC Hz Hz Gas Requirements O 2 1 bar 2 bar CO 2 1 bar 2 bar N 2 N/A 2 bar Air 1 bar 2 bar System Specifications Dimensions (mm) Docking Station 200 x 120 x 200 Fully-integrated device Controller 260 x 300 x x 1700 x 890 Perfusion 200 x 120 x 200 Weight Docking Station 2.5 kg ~ 300 kg Controller 3.5 kg Instrumentation ph probes 1 (classical) 2 (single-use) D.O. probes 1 (classical) 2 (single-use) Temperature probes 1 2 Biomass probes 1 (classical) 1* (single-use) (single-use) Pumps 1 for NaOH addition 5 Perf IN/OUT, NaOH Perfusion module (optional) addition,sampling and accessories Control System Architecture PC-HMI PLC-SCADA Compliance GAMP & 21 CFR Part 11 GAMP & 21 CFR Part 11 compliant (optional) compliant (optional)
4 Startup Requirements ph Range Detection range = Detection range = ph 0 14 ph Precision = +/ ph 7.0 Regulation NaOH/CO 2 addition NaOH/CO 2 addition D.O. Range 0-100% 0-100% Regulation O 2 addition Air/O 2 /N 2 additions Temperature Range 15 to 40 C 25 to 40 C Regulation Peltier elements Double jacket Cryostat Speed Range 0-1,500 rpm rpm Mixing System Magnetic drive impeller Magnetic drive impeller (integrated) (integrated) Perfusion Regulation External perfusion system Perfusion system (self-regulated pumping integrated outflow - automatic and manual modes) Gas Flow Rates O ml/min 0-7 L/min CO ml/min 0-2 L/min N ml/min 0-1 L/min Air 0-50 ml/min 0-2 L/min Data Recording/ Available on external Optional Trending laptop (optional) Production NA Optional Sequences Mode and Alarm Management System (self-regulated pumping outflow - automatic and manual modes) Data Management with the icellis 500 Bioreactor The icellis 500 bioreactor is operated through a fully-integrated process control unit designed to fit with GAMP and 21 CFR Part 11 requirements. It operates with a SCADA system (intouch) developed by Wonderware which offers maximum functionality for process control and data acquisition. The data and alarms generated during the cell culture batch are stored on a local hard-drive in a secured Wonderware format. An additional network interface card, as well as an Ethernet port, is included in the system to connect it to the user s network and enable custom data history. In addition, the system allows real-time alarming (option) by sending the pre-defined user an SMS or message in case of an unexpected event. Pall Life Sciences IT experts can support integration of the icellis process control system in GMP environments. Performance The fixed-bed design of the icellis system makes it the perfect solution for process intensification, providing a large cell growth surface area while keeping the volumes very small. In only 25 L of volume, icellis accommodates up to 500 m 2 of growth area, which, when compared with available surfaces of multitray systems and roller bottles, corresponds to a huge reduction in volumes. Table 2 illustrates the icellis 500/500 m 2 system is equivalent to 794 cell stacks with 10 trays and 5,882 Roller Bottles at 850 cm 2 each. General overview from icellis 500 software Split view of icellis nano disposable bioreactor with reusable lid Bioreactor overview from icellis 500 software 4
5 Table 2 Available surface area in an icellis 500 system and comparison with multitray cell factory systems (CF)/Roller Bottles (RB). Equivalent CF10 Equivalent RB Equipment Fixed-Bed Volume (L) Culture Surface Area (m 2 ) (6,300 cm 2 ) (850 cm 2 ) icellis nano Fixed-bed height 10 cm icellis 500/ ,882 Fixed-bed height 10 cm In addition to volume reduction, the use of a filled, fixed-bed in the icellis system enables higher specific productivity than in classical systems. To demonstrate the impact of icellis bioreactors on cell densities and productivity, many cultures were performed with several cell lines (Table 3). Results clearly highlight that the growth matrix inside the icellis system is a favored growth environment which, together with controlled cell culture parameters, can significantly increase the biomass amplification, as well as the number of viral particles produced by each cell (specific productivity). Additional experiments demonstrated that icellis bioreactors require limited adaptation protocols and enable easy, straightforward scale-up for the majority of mammalian cell lines. Table 3 Cell densities and specific productivities achieved with icellis systems (several mammalian cell lines). Comparison is with multitray cell factory systems (CF)/Roller Bottles (RB). Cell Densities in Specific Productivity (/cm 2 ) Equivalent CF/RB Equivalent CF/RB Cell Line Bioproduct icellis Bioreactor Control icellis nano 4 m 2 500/500 (500 m 2 ) Vero Serum-Free Paramyxovirus 33.7 x 10 6 cell/ml T-Flask 6.37 x CF10 8,750 CF x 10 4 TCID50 TCID RB 64,625 RB A549 raav 9 x 10 4 cell/cm 2 CS5, 3 x x CF10 1,250 CF10 vg/cm 2 vg/cm 2 75 RB 9,375 RB HEK293 CGTC x 10 5 cell/cm 2 Hyperflask 3.8 x 10 9 TCID50 35 CF102 4,413 CF10 7 x 10 8 TCID50 63 RB 32,946 RB CEF Serum-Free MVA Virus 30 to 100 cell/ml 3.48 x 10 6 PFU 5 CF CF10 (non-lytic infection) 38 RB 4,750 RB MDBK BHV 60 cell/ml 2.17 x CF10 2,000 CF10 TCID RB 15,000 RB VERO Serum-Free Influenza 28 cell/ml 3.75 x CF10 1,875 CF10 TCID RB 13,750 RB VERO Undisclosed 75 to 150 cell/ml Undisclosed Undisclosed 10 CF10 1,238 CF10 Human Vaccine (max) VERO Serum-Free Undisclosed 75 to 150 cell/ml Undisclosed Undisclosed 6 CF CF10 Human Vaccine (max) Corporate Headquarters Port Washington, NY, USA toll free (USA) phone biopharm@pall.com European Headquarters Fribourg, Switzerland +41 (0) phone LifeSciences.EU@pall.com Asia-Pacific Headquarters Singapore phone sgcustomerservice@pall.com Visit us on the Web at us at bioreactors@pall.com International Offices Pall Corporation has offices and plants throughout the world in: Argentina, Australia, Austria, Belgium, Brazil, Canada, China, France, Germany, India, Indonesia, Ireland, Italy, Japan, Korea, Malaysia, New Zealand, Norway, Philippines, Poland, Russia, Singapore, South Africa, Spain, Sweden, Switzerland, Taiwan, Thailand, United Kingdom, and Vietnam. Distributors in all major industrial areas of the world. To locate the Pall office or distributor nearest you, visit The information provided in this literature was reviewed for accuracy at the time of publication. Product data may be subject to change without notice. For current information consult your local Pall distributor or contact Pall directly. 2015, Pall Corporation. Pall,, and icellis are trademarks of Pall Corporation. indicates a trademark registered in the USA and TM indicates a common law trademark. Filtration. Separation. Solution.is a service mark of Pall Corporation. 1/15, PDF, GN USD2941 (1)
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