iCELLis® Bioreactors

Single-use bioreactor for process enhancementHighlights 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 oxygen sparging 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

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OverviewThe 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 thebenefits of a fixed-bed system. The compact system—designedfor quick implementation and ease-of-use—represents a newgeneration of single-use bioreactors.

Central to the iCELLis bioreactor technology is the use of acompact fixed-bed, filled with proprietary macrocarriers. Thismatrix is made of medical grade polyester microfibers and provides – in only 25 L volume – a surface area for cell growthup to 500 m². This is the surface-equivalent of 3,000 RollerBottles (at 1700 cm2 each). The iCELLis bioreactor comes pre-packed with macrocarriers. Compared to a standard stirred-tankbioreactor using microcarriers, the iCELLis technology leads to adramatic simplification of the cell culture process. It alleviatesdelicate and time-consuming manual operations, reducing the overall process time and making the cell culture processmore robust.

Because of the cell to cell interactions within the 3D environmentof the fixed bed, iCELLis bioreactors can be inoculated at verylow density as low as 1,000 cells per cm² (Vero cells). The number and volume of pre-culture steps are therefore reducedconsiderably. As a result of this process simplification, manualoperations and associated costs are greatly reduced.

Evenly-distributed media circulation is achieved by a built-inmagnetic drive impeller, ensuring low shear stress and high cellviability. The cell culture medium flows through the fixed-bedfrom the bottom to the top. At the top, the medium falls as athin film down the outer wall where it takes up O2 to maintainhigh 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 maintainhigh cell densities—equaling the productivity of much largerstirred-tank units.

Process simplification, from vial thawing to final product, is at theheart of the iCELLis platform. As cells are immobilized in thefixed-bed, and the iCELLis system operates in perfusion mode,no centrifugation is needed to harvest the cells. This benefit further simplifies the downstream process.

Features and ConfigurationsIntegrated mixing system for evenly-distributed mediacirculation 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 toensure process reliability

Modular height of fixed-bed – from 2 cm to 10 cm – offeringseveral 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 (up to 4 m²)

The iCELLis 500 system for industrial scale manufacturing (up to 500 m2)

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The iCELLis 500 bioreactor (500 m2) is equivalent to 3,000Roller Bottles (1700 cm2 each)

Microscopy view of MDCKcells immobilized oniCELLis system carriers

Media flow in the iCELLis 500bioreactor (60-70 L)

The iCELLis 500 bioreactor on itsfully-integrated skid

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iCELLis Nano bioreactor

The iCELLis Nano bioreactor is the scale down of the productionscale iCELLis 500 bioreactor. It offers surface for growth from0.53 to 4 m². Both systems use exactly same carriers and samefixed bed heights, providing linear scalability between them. Thisis based on the same principle as chromatography columns.

Figure 1 iCELLis bioreactor bench (Nano) and manufacturing (500) scales and their modular size fixed bed.LC = Low compaction; HC = High compaction.

These features and its low footprint make the iCELLis Nanobioreactor the perfect system for process development or manufacturing of small batches.

A stainless steel lid is screwed on the single-use bioreactor then the whole assembly is autoclaved. It is then placed into its docking station, which is connected to a controller ensuringcontrol, monitoring and recording of all culture parameters, withthe support of a SCADA (supervisory control and data acquisition)system. A perfusion module consisting of 2 pumps is suppliedas an option to manage medium flow to and from the iCELLis Nano bioreactor.

iCELLis 500 bioreactorThe iCELLis 500 bioreactor is the production scale iCELLis system.It offers surface for growth from 66 up to 500 m² in a single-usebioreactor made of material free of animal origin componentsand tested for Class VI USP compliance. The bioreactor is seton a fully integrated system including docking station, perfusionpumps and controller. Coupled to a 21 CFR part 11 compliantsoftware, it will control all process parameters – pH, DO, gasflow, temperature, stirring speed, biomass - throughout theduration of the culture.

As the iCELLis 500 bioreactor fits in a footprint of just 3.2 m², it provides a compact cell culture solution for manufacturing of large batches of clinical material.

Bioreactor Fixed-Bed Fixed Bed Bioreactor Surface Area (m2)Bioreactor Diameter (mm) Height (mm) Volume (L) Volume (L) Low Compaction High Compaction

iCELLis Nano 110 20 0.04 1 0.53 0.8iCELLis Nano 110 40 0.08 1 1.06 1.6iCELLis Nano 110 100 0.2 1 2.6 4iCELLis 500/100 860 20 5 70 66 100iCELLis 500/200 860 40 10 70 133 200iCELLis 500/500 860 100 25 70 333 500

Table 1Configurations of iCELLis bioreactors at small and manufacturing scale

iCELLis Nano

Bench-scale

iCELLis500 Production scale

Fixed Bed Height and/or compaction

increase

4m

LC HC LC HC

2.6m

1.6m 1.08m

0.8m 0.53m

500m 333m

200m 133m

100m 66m

Scale-up Scale-down of diameter

10 cm

4 cm

2 cm

4m2.6m

HCLC

10 c

LC

333m

cm

HC

500m

1.6m

incre

Hcompa

a Heightt and/or Fixed

1.08m

0.8m0.53m

4 c

2 c

133m

ease

r action and/or

Bed66m

m

m

200m

100m

scaleBench-Nano

iCELLis diamof

- ScaleScale

Produc iCEL

meterdown -up e

scalection LLis500

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Startup Requirements

General overview from iCELLis 500 software

Bioreactor overview from iCELLis 500 software

Bioreactor iCELLis Nano iCELLis 500

pH Range Detection range = Detection range =

pH 0 – 14 pH 6 .0– 9.0Precision = +/– 0.01 @ pH 7.0

Regulation Base/CO2 addition Base/CO2 additionD.O.Range 0-150% 0-150%Regulation Air/O2/N2 addition Air/O2/N2 additionTemperature Range 15 to 40 °C 25 to 40 °CRegulation Peltier elements Double jacket – CryostatSpeedRange 0-1,500 rpm 0-400 rpmMixing System Magnetic drive impeller Magnetic drive impeller

(integrated) (integrated)PerfusionRegulation External perfusion system Perfusion system

(self-regulated pumping integrated outflow - automatic and (self-regulated pumping manual modes) outflow - automatic and

manual modes)Gas Flow RatesO2 0-200 mL/min 0-7 L/minCO2 0-10 mL/min 0-2 L/minN2 0-50 mL/min 0-1 L/minAir 0-50 mL/min 0-2 L/minData Recording/ Available on external AvailableTrending laptop (optional)Production NA OptionalSequences Mode and Alarm Management System

Process Control SystemsStartup Requirements

Bioreactor iCELLis Nano iCELLis 500

Power Requirement 500 W 5030 W (230 V) / 4110 W (110 V)

Power Supply 100 – 230 VAC 100 – 230 VAC50-60 Hz 50-60 Hz

Gas RequirementsO2 1 bar 2 barCO2 1 bar 2 barN2 1 bar 2 barAir 1 bar 2 bar

System Specifications

Bioreactor iCELLis Nano iCELLis 500

Dimensions W x H x L(mm)Docking Station 200 x 120 x 200 Fully-integrated deviceController 350 x 350 x 190 1490 x 1670 x 850Perfusion 200 x 120 x 200Weight Docking Station 2.5 kg ~ 485 kgController 8.5 kgInstrumentationpH probes 1 (classical) 2 (single-use)D.O. probes 1 (classical) 2 (single-use)Temperature probes 1 2Biomass probes 1 (classical) 1 (single-use)(single-use)Pumps 1 for base addition 5 – Perf IN/OUT, base

Perfusion module (optional) addition, sampling and accessories

Control SystemArchitecture PC-HMI PLC-SCADACompliance GAMP & 21 CFR Part 11 GAMP & 21 CFR Part 11

compliant (optional) compliant

Data Management with the iCELLis 500BioreactorThe iCELLis 500 bioreactor is operated through a fully-integratedprocess control unit designed to fit with GAMP and 21 CFR Part11 requirements. It operates with a SCADA system (inTouch)developed by Wonderware which offers maximum functionalityfor process control and data acquisition. The data and alarmsgenerated during the cell culture batch are stored on a localhard-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 enablecustom data history. In addition, the system allows real-timealarming (option) by sending the pre-defined user an SMS or e-mail message in case of an unexpected event. Pall LifeSciences IT experts can support integration of the iCELLisprocess control system in GMP environments.

PerformanceThe fixed-bed design of the iCELLis system makes it the perfectsolution for process enhancement, providing a large cell growthsurface area while keeping the volumes very small. In only 25 Lof fixed bed volume, iCELLis accommodates up to 500 m2 ofgrowth area, which, when compared with available surfaces ofmultitray systems and roller bottles, corresponds to a hugereduction in volumes. Table 2 illustrates the iCELLis 500/500 m2

system is equivalent to 794 cell stacks with 10 trays and 5,882Roller Bottles at 850 cm2 each.

Table 2Available surface area in an iCELLis 500 system and comparison with multitray cell factory systems (CF)/Roller Bottles (RB).

Equivalent CF10 Equivalent RBEquipment Fixed-Bed Volume (L) Culture Surface Area (m2) (6,300 cm2) (850 cm2)

iCELLis Nano 0.2 4 6 47Fixed-bed height 10 cmiCELLis 500/500 25 500 794 5,882Fixed-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 classicalsystems. 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 environmentwhich, 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 requirelimited adaptation protocols and enable easy, straightforward scale-up for the majority of mammalian cell lines.

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Table 3Cell 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 Equivalent CF/RB Equivalent CF/RBCell Line Bioproduct iCELLis Bioreactor Control iCELLis Nano 4 m2 500/500 (500 m2)

Vero Serum-Free Paramyxovirus 33.7 x 106 cells/mL T-Flask 6.37 x 105 71 CF10 8,750 CF10 5.67 x 104 TCID50 TCID50 527 RB 64,625 RB

A549 rAAV 9 x 104 cells/cm2 CS5, 3 x 106 5 x 108 10 CF10 1,250 CF10 vg/cm2 vg/cm2 75 RB 9,375 RB

HEK293 CGTC102 1.68 x 105 cells/cm2 Hyperflask 3.8 x 109 TCID50 35 CF102 4,413 CF10 7 x 108 TCID50 63 RB 32,946 RB

CEF Serum-Free MVA Virus 30 to 100 x 106 cells/mL 3.48 x 106 PFU 5 CF10 625 CF10(non-lytic infection) 38 RB 4,750 RB

MDBK BHV 60 x 106 cells/mL 2.17 x 106 16 CF10 2,000 CF10TCID50 120 RB 15,000 RB

VERO Serum-Free Influenza 28 x 106 cells/mL 3.75 x 105 15 CF10 1,875 CF10TCID50 110 RB 13,750 RB

VERO Undisclosed 75 to 150 x 106 cells/mL Undisclosed Undisclosed 10 CF10 1,238 CF10Human Vaccine (max)

VERO Serum-Free Undisclosed 75 to 150 x 106 cells/mL Undisclosed Undisclosed 6 CF10 794 CF10Human Vaccine (max)

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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 yourlocal Pall distributor or contact Pall directly.

© 2016, Pall Corporation. Pall, , and iCELLis are trademarks of Pall Corporation. ® indicates a trademarkregistered in the USA and TM indicates a common law trademark. Filtration. Separation. Solution. is a servicemark of Pall Corporation.

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