SingleSingle--use Process Componentsuse Process Components““DisposablesDisposables””: :

Some Process Engineering considerationsSome Process Engineering considerations

Peter KRAEMER Sanofi AventisIA C&B’s Direction Technique / Engineering

Joint Workshop of ISPE - DACH & France“Single-use Technologies in Biomanufacturing Processes”MarseilleMay 26 & 27, 2010

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“Disposables” (SU-PC) concepts: Scope

Reminder: drivers for alternative technology set-upsProcess Engineering considerations

Process scale / Processing volumes & Mixing characteristicsInstrumentationConstruction material for single-use process itemsHSE considerationsConnecting systems / mechanical connectors / tube weldingStoraging and shipment

Case studies: Cost estimationsConclusions

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“Disposables”:Drivers for alternative technology set-ups

Are you looking for a technology, which is highly flexibledoesn’t need to be suitable for a full chemical / ATEX environment allows accelerated implementationrequires less investmentallows accelerated start-up……

Then, letThen, let’’s check out for Singles check out for Single--use process componentsuse process components((““DisposablesDisposables””) technology !!) technology !!

Con’s & limitations? They are not forgotten – see end of the presentation !

® Peter KRAEMER – sanofi aventis

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“Disposables”: Process Engineering Considerations (1/3)

Process scale / Processing volumes: Transfer timesTransfer times

Tube diameter (internal) Flow rate (l/min) Transfer time (min) (“ / mm) @ 1m/s ∆p = 0,1 barg (@ 100 l / 1.000 l)

¼ / 6,4 1,7 59 / 167 ½ / 14,8 10.3 10 / 68 ¾ / 20,4 19,3 5 / 49 1 / 25,6 30,9 4 / 39

Max. dimension currently available for

validated flange /coupling weldings

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“Disposables”: Process Engineering Considerations (2/3)

Mixing characteristics – comparisons of technologies

Note: Mixing characteristics could be acceptable; but mass transfer requires superior performance !

-;(

-;I

-;)

- limitations

® Peter KRAEMER – sanofi aventis

Turn-over times: min/hr

Hose diameter/pump rate

Turn-over times: ≈ min(s)

Agitator design & speed / pump rate

Turn-over times: < min.

Tilt angle / Mechanical design

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a. XDR™-Disposable Stirred Tank Bioreactor, e.g. XCELLEREXb. S.U.B. Single-Use Bioreactor, e.g. Hyclone-ThermoScientificc. ’Pad-Drive ™ Disposable bioreactor, e.g. Artelis-ATMI Life-Sciencesd. LevTech-System, floating agitator and superconductor drive, Stedim-Sartorius

d

Examples for mixing/agitating systems

“Disposables”:Process Engineering Considerations (3/3)

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“Disposables”: Instrumentation

Possible instrumentationAdapted standard technology

Disposable sensors

Physical parameter

Classical process equipment design

Disposable TechnologySingle use design

L Level, (cont.) Scale / weight, radar Scale, Weight

P Pressure Pressure transmitter Disposable sensor integrated in bag or manifold; coupled to re-usable transmittertransmitter detached to system via membranes

T Temperature Pt 100; Thermometer

Similar sensor technology; placement of probe:- inserted in "pocket"/sheaft; or- access via sterile pipe connector

W Weight Scale, external load cells Scale, external load cells

F Flow Mass flow meters;turbinesrotameter

liquids:- disposable sensor integrated in bag or manifold; coupled to re-usable transmitter- inline measure measurement with flow meters prior to sterile filter- indirect via weight increments (balances)gas: gas flow meters prior to sterile filter

Q pH Conventional electrode Optic or conventional via sterile pipe connector

Q pO2 Conventional electrode Optic or conventional via sterile pipe connector

Q Conductivity Conventional electrode Conventional via sterile pipe connector

S Frequency (agitator)

conventional conventional (installed on drive)

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“Disposables”:Stainless steel or disposable components

Potential for displacement of disposable technology vs. stainless steel components?

® Peter KRAEMER – sanofi aventis

Components in contact with product solutions:wide range of disposable applications

Support devices / construction: still in stainless due to mechanical strength, and …- up to 200 mUS$ for bioreactor platform (2 m ³ scale)- up to 30 mUS$ for bag stands (2 – 3 m³ scale)- up to 30 mUS$ for agitated tanks (+ 10 mUS$ for load cells)

√

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Formulation / Filling / shipment

• LevTech mixing bag system• no SIP or CIP piping required

Formulation and Fill Equipment Controlled freezing: scope of (auxiliary) equipment

Storage & Shipment• Bags or tanks

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Bags: Bioproduct storage & shipmentControlled freezing/thawing options

Idea and reality: required auxiliary equipment

Transfer carts:Different bag sizesFacilitate transport

Storage moduless:100 l capacityHigh density storage in cold roomDetachable trolley

Filling station:Facilitates the filling procedureMinimizes container handling

Shippers:Ensure thermal stability during shipping

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Bags: Bioproduct storage & shipmentControlled freezing/thawing options

Idea and reality: shipping containersShips 16,6 l or 8,3 l bagsRigid plastic shell, closed cell polyurethane foam interior, foam bumpersRefrigerant demand: approx. 36 kg dry-iceTotal laden weight approx. 90 kgThermal testing:

External: + 20 to + 40 °CCELSIUS-Pak internal: < - 40 °C for 120 hrs. (5 days)

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Bags: Bioproduct storage & shipmentControlled freezing/thawing options

Idea and reality: Bag damages (loose connectors, ice drops)At the beginning: ≈ 10 % of shipped bags leakingAfter thawing unit implementation: ≈ 5 % leaking bagsTarget after improving the full scope of involved items: ≈ 1%

Economic impact:16,6 l @ 40 – 60 g/l ≈ 700 g / bag@ 500 $ / g 350 m$ / bag15 – 20 bags / batch

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“Disposables”: Case studies - Opportunities USP

Production approachFed batch cultivation

Process steps

1 1.000 ml shake flask and Media Preparation 5 ltr Bioreactor

220 ltr BioreactorCultivation time: up to 4 days Feeding Vessels

3100 ltr BioreactorCultivation time: up to 4 days

4500 ltr BioreactorCultivation time: up to 4 days

52,5 m³ BioreactorCultivation time: up to 14 days

Utilities6 Pure steam

Disk Stack Centrifuge WFIDepth filtration Chilled waterPolishing Filter 0,22 µm CIP unit

Air, CO2, O2, N2

Inactivation7

2,5 m³ SS vessel

BD CapI ProcessCultivation and Harvesting

Main Process Equipment

10 batches / yearBioreactor productivity up tp 5 g/ liter

DownstreamProcessing

Inoculation lab

Precultivation

Maincultivation

Isolation

Harvesting

Thawing & Expansion 1 & 2

Expansion 3

Harvesting

Cells Separation

Expansion 4

Expansion 5

Production

Production approachFed batch cultivation

Process steps

1 1.000 ml shake flask and Media Preparation 5 ltr Wave bag

220 ltr Wave bagCultivation time: up to 4 days

3100 ltr Wave bag or100 l Disposable BioreactorCultivation time: up to 4 days Media Storage

4500 ltr Disposable BioreactorCultivation time: up to 4 days

52,5 m³ BioreactorCultivation time: up to 14 days

UtilitiesPure steam

6 WFIDisk Stack Centrifuge Chilled wateror disposable centrifuge CIP unitDepth filtration Air, CO2, O2, N2

Polishing Filter 0,22 µm InactivationDecon Autoclave

73 x 1 m³ Disposable bags

BD CapI ProcessCultivation and Harvesting

Main Process Equipment

10 batches / yearBioreactor productivity up tp 5 g/ liter

DownstreamProcessing

Inoculation lab

Pre Cultivation

Main Cultivation

Isolation

Harvesting

Thawing & Expansion 1 & 2

Expansion 3

Harvesting

Cells Separation

Expansion 4

Expansion 5

Production

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“Disposables”: Case studies - Opportunities DSP

Production approachFed batch Cultivation

Process stepsBuffer Preparation

8 Buffer Storage

Column, Elution and Pool VesselVirus Inactivation reactor

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Column, Elution and Pool Vessel

11 UtilitiesPure steam

Column and Pool vessel WFIChilled water

CIP unit12 Cold Storage

Filter and Pool vessel

13Ultra/Diafiltration

Freezing14

10 Batches / yearOverall Yield 75 %

Main Process Equipment

BD CapI ProcessDownstream Processing

Hydrophobic-InteractionChromatography

Viral Clearance Filtration

Concentration

Final filling of API

Ion Exchange Chromatography

Affinity Chromatography

Production approachFed batch Cultivation

Process stepsBuffer Preparation

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Column, Elution and Pool VesselVirus Inactivation reactor Buffer Storage

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Column, Elution and Pool Vessel

11 Sampling

Column and Pool vessel

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UtilitiesFilter and Pool vessel Pure steam

WFIChilled water

CIP unit13 Cold storage

Ultra/Diafiltration

14 Freezing

BD CapI ProcessDownstream Processing

UpstreamProcessing

10 Batches / yearOverall Yield 75 %

Main Process Equipment

Hydrophobic-InteractionChromatography

Viral Clearance Filtration

Concentration

Final filling of API

Ion Exchange Chromatography

Affinity Chromatography

® Peter KRAEMER / Robert MUELLER– sanofi aventis

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“Disposables”:Cost comparisons – scope of examples

2,5 m³ (wv) fed-batchPilot plant1 DSP train

a) Stainless steel5, 20, 100, 500 & 2.500 l1 DSP trainbuffer / media prep in SS

b) Hybrid design - “mixed” approach because of the 2,5 m³ (wv) of the bioreactor (not available as single use equipment for the time being).- Seed train in wave reactors- 1 DSP train- buffer / media prep as SU components- Utility supply & CIP/SIP to be realized according to the main bioreactor / buffer tank volumes

2 m³ overall capacityCommercial process 1 DSP train

a) Stainless steel - 1 seed train (10 l, 50 l, 200 l) - 2 main reactors @ 1 m³- 1 DSP train (3g/l, 48 batches/a)

b) Single-Use / Disposable - seed train identical steps, - main reactor @ 4 x 500 l disposable wave bioreactors;- media and buffer preparation still in SS

1 m³ perfusionCommercial process SU-PC maximized

Project 1

Stainless Hybride

Project 2

Stainless Single-Use

Project 3

Stainless Single-Use

® Peter KRAEMER / Robert MUELLER– sanofi aventis

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“Disposables”:Economic evaluation of technologies

Investment considerations on referenced projectsInvestment considerations on referenced projects

® Peter KRAEMER / Robert MUELLER– sanofi aventis

Equipment: 25 % Equipment: 25 % –– 45 %, 45 %, depending whether all components can be realized in SU technology, or mix is necessary – see also piping & instrumentation

Piping: 37 % Piping: 37 % –– 45 %,45 %,depending on extent of absence of CIP & SIP pipe-work

Instrumentation & electrical: 36 % Instrumentation & electrical: 36 % –– 52 %, 52 %, depending on extent of absence of CIP & SIP; automated valves, less automation, basic process control system approach

Building (incl. HVAC & Labs): 0 % Building (incl. HVAC & Labs): 0 % –– 22 %;22 %;almost no saving expected - literature data are more optimistic

Engineering: 24 % Engineering: 24 % –– 40 %; 40 %; depending extent of SU technology with impact on piping & instrumentation

Overall: 24 % Overall: 24 % -- 40 % lower investment in SU technology40 % lower investment in SU technology

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“Disposables”:Economic evaluation of technologies

Impact of extent of facility utilization on Opex at 2 – 3 m³ scale(10 - 20 - 40 batches per year; 1st year of operation)

k€ k€ k€

® Peter KRAEMER / Robert MUELLER– sanofi aventis

Consumables / incl. DisposablesConsumables / incl. Disposables: 86 % - 102 % increase in case of SUResinsResins: similar cost for both approaches; scale is regarded beyond SUEnergy & Utilities:Energy & Utilities: 8 % - 25 % savings in case of SU (mainly CIP, PW / WfI)MaintenanceMaintenance: 20 % – 25 % savings for SULabor:Labor: 12 % - 13 % increase in case of SU Waste:Waste: impact negligible at industrial site ! (might look different for R&D environment with external incineration)Depreciation:Depreciation: 20 % savings in case of SU

Overall: 0 % Overall: 0 % -- 30 % higher Opex for SU with increasing batch #30 % higher Opex for SU with increasing batch #

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“Disposables”: Technology score card (1/2)

® Peter KRAEMER / Robert MUELLER– sanofi aventis

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“Disposables”:Technology score card (2/2)

® Peter KRAEMER / Robert MUELLER– sanofi aventis

Benefits & limitations: Application and scale are key drivers !

Flexibility and time for implementation: Clear advantages for SUs !

Regular manufacturing at “larger” scale: stainless steel still competitive !

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“Disposables”: Application options

Technology preference per develpment phase

0,0

0,1

1,0

10,0

100,0

1.000,0

10.000,0

100.000,0

1.000.000,0B

iore

acto

r vol

ume

(l)

ScreeningDevelopment

ProcessDevelopment

Clinical materialSmall/large scale

Production

10-2

101

100

10-1

103

102

104

105

105

Single-UseProcess style

Hybrid design

Stainless steel

Single-UseLab style

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“Disposables”: Contributors

Special thanks:Special thanks:Robert MUELLER

Dieter BECKERHelmut MEIERCarsten MUELLERRolf NONNENMACHERHolger GACHOTMichael VOSS

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Thanks for your attention!

“Disposables”: Opportunities & constraints