icellis Single-Use Fixed-Bed Bioreactor System

Transcription

1 USD3262 icellis Single-Use Fixed-Bed Bioreactor System Prodctivity and redced footprint for virs prodction in adherent cell growth The icellis bioreactor system is an atomated, single-se, fixed-bed bioreactor, that provides excellent cell growth conditions for adherent cells. The bioreactors system main featres inclde: Cell cltre process simplification, from vial to prodct Flly-integrated, single-se bioreactor with disposable, pre-installed calibrated probes Uniqe waterfall system for optimal oxygenation and CO2 stripping Straightforward implementation for faster process development Predictable scalability from R&D to manfactring ompact high-cell-density, fixed-bed bioreactor providing a significant increase in volmetric C prodctivity vs. traditional stirred tanks Significant decrease in operational costs and capital investments vs. classical 2D flatware

2 Central to the icellis bioreactor technology is the se of a compact fixed-bed, filled with proprietary macrocarriers. This matrix is made of class VI (1) polyester microfibers and provides in only 25 L volme a srface area for cell growth p to 500 m 2. This is the srface-eqivalent of 3,000 roller bottles (at 1700 cm² each) (Figre 1). The icellis bioreactor comes pre-packed with macrocarriers. Figre 2 Media flow in the icellis 500 bioreactor (60-70 L) Becase of the cell to cell interactions within the 3D environment of the fixed-bed, icellis bioreactors can be inoclated at very low density, 3,000 cells per cm 2 and even lower (VERO cells). The seed train can as sch be streamlined and simplified, leading to less manal operations and a redction of associated costs. Evenly-distribted media circlation is achieved by a bilt-in magnetic drive impeller, ensring low shear stress and high cell viability (Figre 2). The cell cltre medim flows throgh the fixed-bed from the bottom to the top. At the top, the medim falls as a thin film down the oter wall where it takes p O 2 to maintain high k L a in the bioreactor. This niqe waterfall oxygenation, together with a gentle agitation and biomass immobilization, enables the compact icellis system to achieve and maintain high cell densities eqaling the prodctivity of mch larger stirred-tank nits. Figre 3 Microscopy view of MDCK cells immobilized on icellis system carriers Process simplification, from vial thawing to final prodct, is at the heart of the icellis platform. As cells are immobilized in the fixed-bed (Figre 3), and the icellis system operates in perfsion mode, no centrifgation is needed to harvest the cells. This benefit frther simplifies the downstream process. Figre 1 The icellis 500 bioreactor (500 m 2 ) is eqivalent to 3,000 roller bottles (1700 cm 2 each) System Overview, Featres and Benefits The icellis bioreactor system is available in two formats (Table 1 and Figre 5): The icellis Nano system for feasibility stdies and small-scale prodction (p to 4 m 2 ) The icellis 500 system for indstrial scale manfactring (p to 500 m²) Table 1 Configrations of icellis bioreactors at small and manfactring scale Bioreactor Fixed-Bed Fixed-Bed Bioreactor Srface Area (m 2 ) Bioreactor Diameter (mm) Height (mm) Volme (L) Volme (L) Low Compaction High Compaction icellis Nano icellis Nano icellis Nano icellis 500/ icellis 500/ icellis 500/

3 Both systems have an integrated mixing system for evenly-distribted media circlation and low shear stress, waterfall media oxygenation for high oxygen transfer and se the same macrocarriers specifically adapted to adherent cell cltres. The height of fixed-bed can be varied from 2 cm to 10 cm offering several configrations of small and large scale. The scaling from the small bioreactor to the large one is straightforward, and typically based on retention of fixed-bed height (Figre 4). Figre 4 icellis bioreactor bench (Nano) and manfactring (500) scales and their modlar size fixed-bed. LC = low compaction; HC = high compaction Figre 5 icellis Nano (left) and icellis 500 (right) Table 2 icellis Nano bioreactor system Featre Benefit Bioreactor with re-sable electrochemical sensors Hardware featres docking station with perfsion integrated and mpath control tower and Link Server IP54 rated hardware Advanced alarming Overflow control and internal bioreactor pressre measrement Temperatre control and advanced heating throgh resistors Gas handling by six oxygen-rated thermal mass flow controllers (TMFCs) Integrated cable management tools mpath control tower designed for operation with Pall Link SCADA software package Monitoring of DO, ph, temperatre and biomass possible Ergonomic and robst indstrial design, sing same controller as all other Pall benchtop bioreactor systems Ready for se in cleanrooms Color coded illmination allows identification of operating state from distance Prevents spill Maintain temperatre and heat from 20 to 37 C within 1 hor Accrate gas mixing enabling accrate parameter control Ease-of-se and ergonomics See Pall pblication USD 3220 for more information Allows access data anywhere on any networked device inclding laboratory workstations, desktop compters or remote devices sch as laptops or mobile phones Intitive visal displays for easy interaction with the bioreactor Advanced trending capability Database centric design spports naming, logging and exporting experiments for traceability and data analysis Management featres for ser-definable recipes inclding schedling, version control and retirement for recipes no longer in se PLC based and designed for operation following 21 CFR Part 11/Annex 11 reglations mpath control tower icellis Nano single-se bioreactor icellis 500 control system icellis 500 single-se bioreactor icellis Nano docking station icellis 500 thermo control nit 3

4 Table 3 icellis 500 bioreactor system Single-se bioreactor made of material free of animal origin components and tested for Class VI USP (1) compliance. The bioreactor is set on a flly integrated system inclding docking station, perfsion pmps and controller. Copled to a 21 CFR Part 11 compliant software, it will control all process parameters ph, dissolved oxygen (DO), gas flow, temperatre, stirring speed, biomass throghot the dration of the cltre. Featre Single-se bioreactor made of USP Class VI (1) materials free of animal origin components and spplied gamma-irradiated Bioreactor with single-se sensors and Pall Kleenpak Hardware featres docking station with perfsion integrated and controller Overflow control and internal bioreactor pressre measrement Temperatre control and heating throgh temperatre control nit and integrated doble jacket Integrated tbing management tools Process control Benefit Ease-of-se, plg and play concept, with no cleaning validation for prodct contact components reqired Monitoring of DO, ph, temp and biomass possible, in addition to bioreactor pressre Ergonomic and robst indstrial design Prevents spill Robst temperatre control Ease-of-se and ergonomics Designed in accordance to Good Atomated Manfactring Practice (GAMP) and for operation following 21 CFR Part 11/Annex 11 reglation SCADA system (intoch ) developed by Wonderware which offers maximm fnctionality for process control and data acqisition The data and alarms generated dring the cell cltre batch are stored on a local hard-drive in a secred Wonderware format An additional network interface card, as well as an ethernet port, is inclded in the system to connect it to the ser s network and enable cstom data history Performance and Applications The fixed-bed design of the icellis system makes it the perfect soltion for indstrialization of adherent cell processes and virs prodction for both gene therapy and vaccines, providing a large cell growth srface area while keeping the footprint very small. In only 25 L of fixed-bed volme, the icellis system accommodates p to 500 m 2 of growth area, which, when compared with available srfaces of mltitray systems and roller bottles, corresponds to a hge redction in volmes. Table 4 illstrates the icellis 500/500 m 2 system is eqivalent to 794 cell stacks with 10 trays and 5,882 roller bottles at 850 cm 2 each. Table 4 Available srface area in an icellis 500 system and comparison with mltitray cell factory systems (CF)/roller bottles (RB) Eqivalent Eqivalent Eqivalent Fixed-Bed Cltre CF10 Roller Bottles Volme (L) Srface (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 volme redction, the se of a filled, fixed-bed in the icellis system enables higher specific prodctivity than in other cltre systems. Table 5 below shows data on yield achieved for most common vectors. Reslts clearly highlight that the growth matrix inside the icellis system is a favored growth environment which, together with controlled cell cltre parameters, can significantly increase the biomass amplification, as well as the nmber of viral particles prodced by each cell (specific prodctivity). Additional experiments demonstrated that icellis bioreactors reqire limited adaptation protocols and enable easy, straightforward scale-p for the majority of mammalian cell lines. Table 5 Specific prodctivities achieved with icellis systems with several kinds of vectors prodced from different mammalian cell lines Extrapolated Vector Cell Size Yield/cm 2 Unit to 500 m 2 Sorce Adeno-Associated HEK 0.53 m x VG 2.3 x University of Ulm Virs (AAV) 293T (transient) Biotechnology Jornal 2015 HEK 0.53 m x VG 1.8 x St Jde 293T/17 (transient) Hman Gene Therapy Methods 2016 Lentivirs HEK 0.53 m 2 5.1x 10 6 TU 2.5 x MolMed 293T (transient) Vector manfactring prodction sing disposable fixed-bed bioreactors icellis Nano. Cell and Gene Therapy Manfactring conference, 2016 Retrovirs AM m x 10 5 TU 3.6 x MolMed (stable) Vector manfactring prodction sing disposable fixed-bed bioreactors icellis Nano. Cell and Gene Therapy Manfactring conference, 2016 HEK 2.65 m x 10 7 VT 4.6 x Memorial Sloan Kettering 293Vec (stable) Jornal of Immnotherapy 2015 Adenovirs HEK 100 m 2 6.1x 10 9 VP 3.0 x FinVector 293 (infection) Hman Gene Therapy 2015 VG = viral genomes, TU = transdcing nits, VT = viral titer, VP = viral particles 4

5 Docmentation and Certificates The icellis bioreactors are delivered with complete docmentation that qalifies the system. All nits are tested prior to shipment and witnessed factory acceptance testing (FAT) is available on reqest. The biocontainers are delivered with all the batch specific certificates and calibration information for the inclded single-se sensors. Pall also can assist with hardware qalification, and standard installation verification (IV), operational verification (OV), and site acceptance test (SAT) test protocols are available. Preventive Maintenance and Service Packages The icellis systems are spported with extensive maintenance and service packages. Pall has a global network of technicians and engineers available to assist with installation, qalification, training, technical spport, preventive maintenance and on-site repairs. Contact Pall to discss any of these services. Pall Accelerator process development services provide cstomers with expert advice and spport for the implementation of flly integrated single-se processing soltions into new or existing processes, inclding process optimization, eqipment selection, transfer of existing processes, scale-p and trobleshooting advice. Details can be fond in the Accelerator Process Development Services brochre (USD 3079). Preventive maintenance packages are available to ensre the continos reliable operation of the icellis bioreactor and to minimize downtime. Preventive maintenance typically incldes a fll fnctional test, exchange of wear parts, and calibration of all sensors. Details can be fond in Maintenance Services brochre (USD 3078). General System Specifications icellis Nano System icellis 500 System Dimensions (W x D x H) (mm) Controller 230 x 600 x x 855 x 1670 Docking station 340 x 360 x 290 Weight (kg) Controller 20 kg 485 kg (withot TCU) Docking station 6.7 kg Gases Connections Qick connectors 6 mm Control TMFC TMFC O ml/min ml/min CO ml/min ml/min N ml/min ml/min Air ml/min ml/min Pmps 1 x fixed speed 5 x (perfsion in-ot, base (base addition); addition, sampling and 2 x variable speed inoclation) (perfsion in-ot) Agitation control Magnetic drive impeller Magnetic drive impeller and range ( rpm) (0-450 rpm) Temperatre control Resistors Peltier Doble jacket TCU and range elements C C ph control Electrochemical, re-se Single-se, optical DO control Electrochemical, re-se Single-se, optical 0-150% air satration 0-150% air satration Control architectre PLC PLC SCADA Pall Link any device Wonderware (local) connected BioXPert W7 (remote) Ordering Information icellis Nano hardware icellis Nano bioreactor system Pall Link server and SCADA software Part Nmber ICLNANOBRS MPATHLINK icellis Nano single-se bioreactors Part Nmber Srface Area for Part Nmber (Biomass Compatible) Cell Growth (m²) NS NS NS NS NS NS NS NS NS NS NS NS 4.00 icellis Nano single-se manifolds Part Nmber M Base manifold (gamma irradiated) N Sampling manifold 1 L (gamma irradiated manifold for sampling, feeding, harvesting) U Recirclation loop manifold 2 L (gamma irradiated) V Lid tbing (non sterile 6 tbing with CPC and vent filter for bioreactor assembly atoclaving) E Recirclation loop manifold 10 L (gamma irradiated) icellis 500 hardware Part Nmber ICL500CS110 ICL500CS230 icellis 500 bioreactor system 110 V icellis 500 bioreactor system 230 V icellis 500 single-se (SU) bioreactors 10 cm falling film Part Nmber 4415-I500V I500V I500V I500V I500V I500V I500V133BM 4415-I500V333BM 4415-I500V100BM 4415-I500V200BM 4415-I500V500BM 4415-I500V66BM 4415-I500V50060 icellis 500 SU vessel 66 m² with Kleenpak icellis 500 SU vessel 133 m² with Kleenpak icellis 500 SU vessel 333 m² with Kleenpak icellis 500 SU vessel 100 m² with Kleenpak icellis 500 SU vessel 200 m² with Kleenpak icellis 500 SU vessel 500 m² with Kleenpak icellis 500 SU vessel 133 m² with Kleenpak and biomass probe icellis 500 SU vessel 333 m² with Kleenpak and biomass probe icellis 500 SU vessel 100 m² with Kleenpak and biomass probe icellis 500 SU vessel 200 m² with Kleenpak and biomass probe icellis 500 SU vessel 500 m² with Kleenpak and biomass probe icellis 500 SU vessel 66 m² with Kleenpak and biomass probe icellis 500 SU vessel 500 m² 6CMFF K3 S All icellis 500 single-se bioreactors exist in 6 cm falling film version contact yor Pall representative for references. 5

6 icellis 500 single-se manifolds Part Nmber 6415-I500MFLA Contains icellis 500 starter kit low flow Allegro film S icellis 500 manifold inoclm 2 and 10 L 1 biocontainer T icellis 500 manifold base addition F icellis 500 manifold feed in 2 low flow G icellis 500 manifold harvest and feed ot 1 low flow V icellis 500 manifold feed in 1 for heating 1 modle low flow U icellis 500 manifold sampling with 1 Kleenpak sterile disconnector Part Nmber 6415-I500MFHA icellis 500 starter kit high flow Allegro film icellis 500 starter kit low flow Allegro film contains Z icellis 500 manifold harvest and feed ot high flow S icellis 500 manifold inoclm 2 L and 1 10 L biocontainer T icellis 500 manifold base addition C icellis 500 manifold feed in 2 high flow R icellis 500 manifold feed in 1 for heating 1 modle high flow U icellis 500 manifold sampling with 1 Kleenpak sterile disconnector Qty Qty The icellis bioreactors are available as standard models and both the bioreactor hardware and consmables can be cstomized. Please contact a Pall representative to find the ideal soltion for yor application. Or teams are specialized in pstream and downstream processing and will gladly help find the right technology for any part of yor manfactring process. It is possible to combine prodcts to meet any reqirement in the pstream part of the process: Media preparation Bffer preparation Media sterilization and aseptic transfer liqid transfer into the bioreactor (e.g. glcose, anti-foam, base, etc.) Seed train soltions Cell harvest and separation Please contact Pall for a total soltion discssion on yor process. (1) As per standards USP <87> Biological Reactivity Test, In Vitro, Cytotoxicity; USP <88> Biological Reactivity Tests In Vivo, for Class VI - 50 C Plastics; and USP <661> Containers - Plastic. Figre 6 icellis Nano bioreactor and mpath control tower Visit s on the Web at s at biotech@pall.com Corporate Headqarters Port Washington, NY, USA toll free (USA) phone biotech@pall.com Eropean Headqarters Friborg, Switzerland +41 (0) phone biotech@pall.com Asia-Pacific Headqarters Singapore phone sgcstomerservice@pall.com International Offices Pall Corporation has offices and plants throghot the world in: Argentina, Astralia, Astria, Belgim, Brazil, Canada, China, France, Germany, India, Indonesia, Ireland, Italy, Japan, Korea, Malaysia, New Zealand, Norway, Philippines, Poland, Rssia, Singapore, Soth Africa, Spain, Sweden, Switzerland, Taiwan, Thailand, United Kingdom, and Vietnam. Distribtors in all major indstrial areas of the world. To locate the Pall office or distribtor nearest yo, visit The information provided in this literatre was reviewed for accracy at the time of pblication. Prodct data may be sbject to change withot notice. For crrent information conslt yor local Pall distribtor or contact Pall directly. 2018, Pall Corporation. Pall,, Allegro, icellis, Kleenpak and mpath are trademarks of Pall Corporation. indicates a trademark registered in the USA. TM indicates a common law trademark. Filtration.Separation.Soltion. is a service mark of Pall Corporation. intoch and Wonderware are trademarks of Schneider Electric Software; Molmed is a trademark of Molmed s.p.a. 2/18, PDF, GN USD3262