BioInnovation Leaders Summit London, February 2015 "Integrated approach to online process monitoring in Cell Culture applications" F. Wolpers Hamilton Bonaduz AG The Heart of Analytics

1. BioInnovationLeadersSummit
London,February2015
Integratedapproachtoonlineprocess
monitoringinCellCultureapplications
F.Wolpers
HamiltonBonaduzAG
TheHeartofAnalytics

2. 2
Chapters
1. ProcessControlChallanges
2. ArcSensors
3. CellDensitySensors
4. SingleUseSolutions
5. Summary
6. AboutHamilton

3. 3
Fermentation–thetraditionalapproach
UseNutrients+MicrobesàMix&Wait=Fermentation
Isyourapplicationmuchmorecomplex?

4. 4
Controliscriticalforfermentation
Nodirectcontrolforcellsavailable:
u Cellstatusmostcritical
u Mainlyofflinemethodsused
u Directmeasurementneeded
u Laborintensive
u Finalresultsavailableatendofprocess
u Notcontinuouscontrol,justasamplerate
Mostonlinemeasurementsareinsufficient:
u pHandDO
u Analogsignalsareweakandunstable
u Sensorlifetimeandmaintenanceiscritical
u notnecessarilyconnectedtothecellstatus

5. 5
OfflineMethodsarepronetoerror
Takeintoconsideration:
u Sampleratefrequencyenough?
u Missingimportantevents?
u Processresultsavailableonlyupto24haftersampling
u Toolatetointerfere
u Lossoftimeandworkincaseoffailure
u Checkgrowthrate
u Analyzeprocessdeviation
u Riskofsterilitybreach(reactorandsample)

6. 6
ArcSensors-integratedonlineprocesscontrol
Arc-IntelligentProcessSensors
Integratedsensorintelligenceeliminates
theneedforexternaltransmitters
u Fullycompensated,digitalsignalà
Improvedrobustnessandreliabilityof
signal
u Simplifiedtroubleshooting
andbuilt-indiagnostics
u In-labcalibration
u Reducesetupcosts
u Easytointegrate
u Validationfriendly:
Documentationofsensor
usehistory

7. 7
ARCConceptOverview
ConventionalSensorSetup
nA,mVanalogue4-20mA
ARCSensorSetup
Modbus,4-20mA
DIGITAL

8. 8
ArcProducts:pH
DifferentapplicationsrequiredifferentpHsensors
► EasyFermPlus
► EasyFermBio
► PolilytePlus
► pHeasy

9. 9
ArcProducts:OpticalDOmeasurement
VisiFerm/VisiProDO:provenmethodfor
measurementofoxygeninprocess
u Simplemaintenancewithnoelectrolyte
u Ruggedsensorcapinfourversionsfor
differentapplications
u NointerferencewithCO2
u Nooxygenconsumption
u Integratedintelligence
u Nopolarization
u Outputsignal:ECS,4-20mA,Modbusor
HART(inVisiProDO)
u Standard12mmformat

10. 10
OnlineMonitoringofViableCellDensity(VCD)
u Capacitancebasedmeasurement
u Accurateandprecisemethodtodeterminelivingcell
concentrations
u Directcorrelationofthebiomasswithprocess
controllingparameterslikepHandDO
u Linkedtobiovolume,
intracellularconductivity,
membranecapacitanceand
cellsizedistribution
u Themeasuringlineconsistsof
capacitancesensor,pre-
amplifierandtransmitter

11. 11
Gainingdirectinformation
ByusingcapacitancebasedBiomassSensors:
u Viablecellsreactascapacitorinalternatingelectricalfield
V~
deadcellsnotpolarized
viablecellspolarized
sensor-tipPlatinumelectrodes

12. 12
Donotmiss“hidden”events
Identifyevents:
u Eventjustbyonlinemeasurementdetected
u Nodetectionwhencountingtotalcells
u Impactonviablecells
time(h)
050100150200250
permittivity (pF/cm)
0
1
2
3
4
5
6
7
8
viable cell count (c/ml)
total cell count (c/ml)
0.0
1.0e+6
2.0e+6
3.0e+6
4.0e+6
5.0e+6
6.0e+6
7.0e+6
8.0e+6
9.0e+6
1.0e+7
permittivity2L
viablecellcount(hema)
totalcellcount(hema)
errorbarsindicate+/-1SD
CourtesyofRoche
Decreaseincapacitance
indicatesanutrientdepletion

13. 13
Investigatecellstatus
Clarifyevents:
u Aftereventdetection,correlationwithadditionaloffline
measurementsforclarification
u Nutrientsandby-products
u Optimalknowledgeonprocessneeded
u Nutrientdepletion
CourtesyofRoche

14. 14
IncreaseEfficiency
Nutrientlimitationisreduced
u Triggerfeedingpumpsbasedoncellstatus
u Higherviabilityandelongatedgrowthtimes,leadto
higheryield
time(h)
5075100125
permittivity (pF/cm)
2
3
4
5
6
7
8
9
glutamine concentration (mmol/L)
glucose concentration (g/L)
0
1
2
3
4
5
alanine concentration (mmol/L)
2
3
4
5
6
7
8
permittivity
glucose
alanine
glutamine
=timeoffeedaddition

15. 15
IncreasedProductivity
Optimizationofharvestingtime:
u Cellsstopproducingat«hidden»eventforunknownreason
u Realizedbycontinuouscontrolofcellstatus
u Processtimewasdecreasedbytwodays
u 10%higherproductivity
CourtesyofMerckSerono
Permitivity (pF/cm)

16. 16
pH,DO,VCDandTCDnowcombined!!
IntegrationofArcSensors,byusingtheArcWirelessConverter
NEW

17. 17
ViewandRecordArcSensors
IntegrationofArcSensorsintoArcViewController
DissolvedOxygen
CellDensity
Conductivity/pH
NEW
EasyfermBio(pH)

18. 18
Reliable(!)SingleUseSolutionforDO&pH
Available2015
u ArcModulepluggedonOneFermSUpHorDOElectrode
u ArcWiWirelessAdapterontopofArcModule
u SystempoweredbyPowerCable
u WirelesscommunicationtoPCwithWirelessConverterorArc
ViewHandheld–soonbasedonBluetooth4.0
u DataloggingwithPCandHDMsoftwareorArcViewHandheld
u Configuration,calibration,displaywithPCandHamiltonGMP
PackagesoftwareorArcViewHandheld
ArcWi
Arc
Module
SUpH
Electrode
PCwithHDMsoftwareand
wirelessconverter

19. 19
SingleUseSolutionforCellDensityMonitoring
TheFuture
<Prototype-Status>
u Underdevelopment
u HamiltonProprietaryIPandPatent
u Usesthesametechnologyasthe
conventionalBiomassSensor
u Diskcanbeweldedinanybag
u ProcessandMediacompatible
materials
u Finaldesignstilltobedetermined

20. 20
Summary
IntelligentArcsensorsdoresolveproblemsofanalogsensors
u SignalRobustness
u LongerLifetime
IntelligentArcsensorsdooffermanynewfeaturesandbenefits
u LabCalibration
u PermanentSelf-Diagnostic
u SensorHistory
OnlineCellDensityMonitoringforincreasedproductivityandyields
u Betterunderstandingofprocessdynamics
u Optimizedfeedingstrategy,inoculationandharvestingtime
u FullprocesscontroltogetherwithpH,DO(andConductivityorORP)

21. 21
60YearsHamilton:AGlobalCompany
500Employees800+Employees
Reno,Nevada
u Syringes
u Pipettes
u DilutersandDispensers
u OEMcomponents
u RoboticLiquidHandling
Franklin,Massachusetts
u Automated-20/-80C
freezers
u CryoVialAutomated
DeCapper
u RackRunner
100Employees
Timisoara,Romania
u Syringes
u Lab-TradeProducts
90Employees
SalesandServiceOrganisations
Bonaduz,Switzerland
u Syringes
u LabandProcess
Sensors
u RoboticLiquidHandling
u MedicalVentilators

22. 22
HamiltonBonaduzAG
ViaCrusch8
CH-7402Bonaduz
Switzerland
Tel.:+41586101010
Fax:+41586100010
contact.pa.ch@hamilton.ch
www.hamiltoncompany.com
Thankyouforyourattention