Multiparameter Sensor System for Single-Use Bioreactors

Transcription

1 Multiparameter Sensor System for Single-Use Bioreactors Strategische Übungen (Electro)chemical Sensors ETHZ, Caspar Demuth Institute of Chemistry und Biotechnology Zurich University of Applied Sciences 1

2 Top 10 largest selling pharmaceutical products (2015) Rank Drug Trade name Type Main indications Company Sales (10 9 USD /year) 1Adalimumab Humira Biologic Rheumatoid arthritis AbbVie Inc. 14,012 1,469 Ledipasvir 2sofosbuvir Harvoni Small molecule Hepatitis C Gilead Sciences 13,864 11,737 3Etanercept Enbrel Biologic Rheumatoid arthritis Amgen/Pfizer 8,697 4,009 4Infliximab Remicade Biologic Crohn's Disease Johnson & Johnson 8,355 1,487 5Rituximab Mabthera Rituxan Biologic Lymphoma/Leukemia/A utommune disorders Roche 7,115 1,456 6Insulin glargine Lantus Biologic Diabetes mellitus Sanofi 7, Bevacizumab Avastin Biologic Metastatic cancers Roche 6, Trastuzumab Herceptin Biologic Breast cancer Roche 6, Lenalidomide Revlimid Small molecule Multiple myeloma Celgene 5, Sofosbuvir Sovaldi Small molecule Hepatitis C Gilead Sciences 5,276 (5,007) vs C. Demuth Strategische Übungen ETHZ

3 Spot the differences! C. Demuth Strategische Übungen ETHZ

4 Spot the differences! Stainless steel bioreactor Single use bioreactor ZHAW ZHAW C. Demuth Strategische Übungen ETHZ

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

6 Relevance of critical bioprocess parameters ph Optimum ph guarantees efficient productivity and high yield Dissolved oxygen (po 2 ) Respiration! Essential in aerobic processes Glucose Main substrate in biotechnological cultivations Lactate and ammonium Metabolites, harmful or toxic to cells µ max ph Maximum specific growth rate (μ max ) of E. coli as a function of ph S.J. Pirt, Principles of microbe and cell cultivation, Blackwell, Oxford, 1975 C. Demuth Strategische Übungen ETHZ

7 Sensor specifications in bioprocesses Wetted parts Compatibility with process Process conditions (p, T, t, SIP, ) Biocompatibility, leachables & extractables Integration Ports, software, protocols Analytical specifications Comparability, robustness, accuracy, Costs «Total cost of ownership» Quality Procedures, approvals, validation, safety C. Demuth Strategische Übungen ETHZ

8 Sensor requirements in single-use reactors Wetted parts Compatibility with process «mild» conditions (gamma irradiation) Use of heat labile materials (polymers, Integration Sensor ist is part of reactor; different ports (no standard) Analytical specifications Comparability, robustness, accuracy, Costs Single Use Quality Different handling and procedures C. Demuth Strategische Übungen ETHZ

9 Selected requirements in single-use bioreactors Process temperature Typically 37 C Process pressure Typically 1 bar Process duration Days to weeks Sterilization procedure Gamma irradiation (25 kgy) Shelf life 1 year Calibration Factory precalibrated, calibration interval > 1 d Measurement range ph po 2 Glucose Lactate Ammonium mbar gl gl gl -1 Response time Seconds Minutes Minutes Minutes Minutes Critical interferences Alkali ions Reducing agents Reducing agents K + C. Demuth Strategische Übungen ETHZ

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 Potentiometric sensors Glass membrane ph Liquid membrane ISE ph, NH 4 + Severinghaus type NH 4 + Amperometric sensors Clark electrode po 2 Enzymatic biosensor Glucose, lactate Optical sensors Fluorescence/absorbance of ph sensitive dye ph Luminescence quenching po 2 Severinghaus optical NH + 4 Ion exchange optode NH + 4 C. Demuth Strategische Übungen ETHZ

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 InSUS 307, Mettler Toledo AG / Thermo Scientifc Ammonium sensors: measurement range and selectivity (nonactin-based sensors) Enyzmatic biosensors: drift and long term stability Biocompatibility of selected materials Nonactin C. Demuth Strategische Übungen ETHZ

12 Proposed sensor system Other solution are feasible as well! Measurement principle ph po2 Glucose Lactate Ammonium optical optical enzymatic biosensor, amperometric enzymatic biosensor, amperometric optical Process temperature Process pressure Process duration ( ) ( ) ( ) Sterilization procedure Shelf life Calibration Measurement range ( ) Response time Interferences C. Demuth Strategische Übungen ETHZ

13 Optical sensors for ph and po 2 X.D. Wang et al, Chem Soc Rev 2014, 43, B.J. Müller et al., Sensors and Actuators B 216 (2015) S.M. Borisov et al, Analyst 2010, 135, C. Demuth Strategische Übungen ETHZ

14 Outlook: Parameters in bioprocess control «readyness» p T physical conductivity po 2 ph pco 2 metabolites sustrates biomass chemical cell size & products morphology biological «relevance» C. Demuth Strategische Übungen ETHZ

15 Process Analytical Technology C. Demuth Strategische Übungen ETHZ

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, C. Demuth, Chem. Unserer Zeit, 2014, 48, C. Demuth Strategische Übungen ETHZ

17 Optical measurements in single-use bioreactors Combination of different measuring modes - Optodes - Fluorescence, Raman - Absorption (UV/VIS) - ATR (IR) Standardized in SU bioreactors needed C. Busse et al., Eng. Life Sci. 2017, 17, C. Demuth Strategische Übungen ETHZ

18 Sensors for online measurement of biomass concentration Dielectric or impedance spectroscopy (commercially available) Online measurement in root cultures J. Carvell at al, BioProcess International 14(3) 2016, C. Demuth et al., Heiligenstädter Kolloquium 2016 C. Demuth Strategische Übungen ETHZ

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, R.A. Potyrailo et al., 2011 IEEE International Conference on RFID-Technologies and Applications C. Demuth Strategische Übungen ETHZ

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

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 C. Demuth Strategische Übungen ETHZ