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Multiparameter Sensor System for Single-UseBioreactorsStrategische Übungen – (Electro)chemical Sensors
ETHZ, 23.10.2018
Caspar DemuthInstitute of Chemistry und BiotechnologyZurich University of Applied Sciences
1
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 2
Top 10 largest selling pharmaceutical products (2015)
https://en.wikipedia.org/wiki/List_of_largest_selling_pharmaceutical_products
Rank Drug Trade name Type Main indications CompanySales (109USD /year)
∆ vs 2014
1Adalimumab Humira Biologic Rheumatoid arthritis AbbVie Inc. 14,012 1,469
2Ledipasvirsofosbuvir Harvoni
Small molecule Hepatitis C Gilead Sciences 13,864 11,737
3Etanercept Enbrel Biologic Rheumatoid arthritis Amgen/Pfizer 8,697 4,0094Infliximab Remicade Biologic Crohn's Disease Johnson & Johnson 8,355 1,487
5RituximabMabtheraRituxan Biologic
Lymphoma/Leukemia/Autommune disorders Roche 7,115 1,456
6Insulin glargine Lantus Biologic Diabetes mellitus Sanofi 7,029 517Bevacizumab Avastin Biologic Metastatic cancers Roche 6,751 2708Trastuzumab Herceptin Biologic Breast cancer Roche 6,603 265
9Lenalidomide RevlimidSmall molecule Multiple myeloma Celgene 5,801 821
10Sofosbuvir SovaldiSmall molecule Hepatitis C Gilead Sciences 5,276 (5,007)
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 3
Spot the differences!
https://www.teachchemistry.org/content/aact/en/periodical/issues/may-2016/chemistry-fun.html
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 4
Spot the differences!
Stainless steel bioreactor
ZHAW
Single use bioreactor
ZHAW
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 5
Task
Propose a multiparameter sensor system for real-time, in situ measurements in single-use bioreactors for critical quantities in bioprocess
control
Include analytical as well as more general aspects
1. Relevance of critical parameters (pH, pO2, glucose, lactate, ammonium)
2. Requirements of a multiparameter sensor system
3. Outline of a sensor system
Measurement principles
Analytical performance vs. requirements
Overall suitability
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 6
Relevance of critical bioprocess parameters
pHOptimum pH guarantees efficient productivity
and high yield
Dissolved oxygen (pO2)Respiration! Essential in aerobic processes
GlucoseMain substrate in biotechnological cultivations
Lactate and ammoniumMetabolites, harmful or toxic to cells
pH
µ max
Maximum specific growth rate (μmax) ofE. coli as a function of pH
S.J. Pirt, Principles of microbe and cell cultivation, Blackwell, Oxford, 1975
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 7
Sensor specifications in bioprocesses
Wetted parts
Biocompatibility,leachables & extractablesCompatibility with
process
Process conditions(p, T, t, SIP,…)
Integration
Ports, software, protocols
Quality
Procedures, approvals,
validation, safetyCosts
«Total cost ofownership»
Analytical specifications
Comparability, robustness, accuracy,…
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 8
Sensor requirements in single-use reactors
Wetted parts
Use of heat labile materials (polymers,
Compatibility withprocess
«mild» conditions(gamma irradiation)
Integration
Sensor ist is part ofreactor; different
ports (no standard)
Quality
Different handlingand procedures
Costs
Single Use
Analytical specifications
Comparability, robustness, accuracy,…
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 9
Selected requirements in single-use bioreactors
Process temperature Typically 37°CProcess pressure Typically 1 barProcess duration Days to weeksSterilization procedure Gamma irradiation (25 kGy)Shelf life 1 yearCalibration Factory precalibrated, calibration interval > 1 d
pH pO2 Glucose Lactate AmmoniumMeasurement range
4-8 0-1000 mbar
0.1-30 gl-1 0.1-10 gl-1 0.01-0.5 gl-1
Response time Seconds Minutes Minutes Minutes Minutes
Criticalinterferences
Alkali ions Reducingagents
Reducingagents
K+
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 10
Available measurement techniques
No single working principle for all parameters combination of electrochemical
and/or optical sensors necessary
A: Electrochemical (potentiometric and amperometric)B: Electrochemical and optical
Potentiometricsensors
Glass membrane pHLiquid membrane ISE pH, NH4+
Severinghaus type NH4+
Amperometricsensors
Clark electrode pO2Enzymatic biosensor Glucose, lactate
Optical sensors Fluorescence/absorbance ofpH sensitive dye
pH
Luminescence quenching pO2Severinghaus optical NH4+
Ion exchange optode NH4+
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 11
Critical aspects
Potentiometric sensors: reference electrode (e.g.
Ag/AgCl)!
pH glass electrode: shelf life, hydration of gel layer
Optical pH sensors: drift, long term stability, bleaching,
dependence on ionic strength
Ammonium sensors: measurement range and
selectivity (nonactin-based sensors)
Enyzmatic biosensors: drift and long term stability
Biocompatibility of selected materials
InSUS 307, Mettler Toledo AG / Thermo Scientifc
Nonactin
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 12
Proposed sensor system
Other solution are feasible as well!
pH pO2 Glucose Lactate Ammonium
Measurementprinciple
optical optical enzymaticbiosensor, amperometric
enzymaticbiosensor, amperometric
optical
Process temperature Process pressure Process duration () () ()Sterilizationprocedure
Shelf life Calibration Measurement range ()Response time Interferences
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 13
Optical sensors for pH and pO2
X.D. Wang et al, Chem Soc Rev 2014, 43, 3666–3761.
B.J. Müller et al., Sensors and Actuators B 216 (2015) 527–534
S.M. Borisov et al, Analyst 2010, 135, 1711-1717.
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 14
Outlook: Parameters in bioprocess control
«relevance»
«rea
dyne
ss»
physical
chemical
biological
pT pH
conductivity pO2pCO2
sustratesmetabolites
biomassproductscell size &
morphology
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 15
Process Analytical Technology
https://www.fda.gov/downloads/drugs/guidances/ucm070305.pdf
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 16
Voltammetric pH measurement
Immobilization of pH sensitive and pH insensitive
redox active components
“calibration free” non glass all solid-state sensor
V.G.H. Lafitte et al, Electrochem Commun 2008, 10,1831–1834
C. Demuth, Chem. Unserer Zeit, 2014, 48, 60 – 67
ww
w.s
enov
asys
tem
s.co
m
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 17
Optical measurements in single-use bioreactors
C. Busse et al., Eng. Life Sci. 2017, 17, 940-952
Combination of different
measuring modes
- Optodes- Fluorescence, Raman- Absorption (UV/VIS)- ATR (IR)
Standardized in SU
bioreactors needed
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 18
Sensors for online measurement of biomass concentration
Dielectric or impedance spectroscopy
(commercially available)
www.aber-instruments.co.uk
J. Carvell at al, BioProcess International 14(3) 2016, 40-48
Online measurement in root cultures
C. Demuth et al., Heiligenstädter Kolloquium 2016
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 19
RFID sensors for SU bioreactors
passive radio frequency identification (RFID) tags on single-use
bioreactors
Sensor antenna changes characteristic resonance, dependent on
physical/chemical properties of process medium
Parameters: pH, conductivity, temperature, biomass,…
R.A. Potyrailo et al, Meas. Sci. Technol. 2011, 22, 082001.
R.A. Potyrailo et al., 2011 IEEE International Conference on RFID-Technologies and Applications
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 20
Sensors for single-use bioreaktors –Challenges
Lifetime Critical for optical pH sensors and biosensors
Recalibration during process (risk of contamination, expenses)
Comparability Comparison of sensors based on different principles
Comparison of online and offline methods is problematic!
Traceable procedures and standards are necessary
Standardization Conventional bioreactors: Standard ports and connectors available
Will allow users to choose best suited sensors for their appilcation
C. Demuth | Strategische Übungen | ETHZ | 23.10.2018 21
Finally…
Single Use: Perfect situation for sensor development growing market
“Disruptive” production technology, new perspectives
Potential for new measurement techniques is not fully exploited yet
Sensor specifications and requirements Not only technical / research viewpoint!
Trends in the process industry favour application of sensors Quality (PAT, QbD)
Automation
Parallelization, Miniaturization
Foliennummer 1Top 10 largest selling pharmaceutical products (2015)Spot the differences!Spot the differences!TaskRelevance of critical bioprocess parametersSensor specifications in bioprocessesSensor requirements in single-use reactorsSelected requirements in single-use bioreactorsAvailable measurement techniquesCritical aspectsProposed sensor system Optical sensors for pH and pO2Outlook: Parameters in bioprocess controlProcess Analytical TechnologyVoltammetric pH measurementOptical measurements in single-use bioreactorsSensors for online measurement of biomass concentrationRFID sensors for SU bioreactorsSensors for single-use bioreaktors – ChallengesFinally…Thank you!Backup SlidesSensoren für Single-Use-BioreaktorenMehrwegsensoren in Einweg-Reaktoren?
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