Online 2D Fluorescence Monitoring in Microtiter Plates Allows Prediction of Cultivation Parameters and Considerable Reduction in Sampling Efforts for Parallel Cultivations of Hansenula polymorpha
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microorganism
2.2. Media Composition
2.3. Precultures
2.4. Main Cultures
2.5. Online Monitoring
2.6. Determination of Offline Parameters
2.7. Spectral Data Processing
2.8. Multivariate Data Analysis
3. Results
3.1. Experimental Layout and Sampling Strategy
3.2. Online and Offline Monitoring of Microtiter Plate Cultivations
3.3. PLS Regression Modelling
3.4. Impact of Numbers of Latent Variables
3.5. Impact of Averaging Spectral Data of Technical Duplicates
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
- Hortsch, R.; Stratmann, A.; Weuster-Botz, D. New milliliter-scale stirred tank bioreactors for the cultivation of mycelium forming microorganisms. Biotechnol. Bioeng. 2010, 106, 443–451. [Google Scholar] [CrossRef] [PubMed]
- Puskeiler, R.; Kaufmann, K.; Weuster-Botz, D. Development, parallelization, and automation of a gas-inducing milliliter-scale bioreactor for high-throughput bioprocess design (HTBD). Biotechnol. Bioeng. 2005, 89, 512–523. [Google Scholar] [CrossRef] [PubMed]
- Szita, N.; Boccazzi, P.; Zhang, Z.; Boyle, P.; Sinskey, A.J.; Jensen, K.F. Development of a multiplexed microbioreactor system for high-throughput bioprocessing. Lab Chip 2005, 5, 819. [Google Scholar] [CrossRef] [PubMed]
- Bolic, A.; Larsson, H.; Hugelier, S.; Eliasson Lantz, A.; Krühne, U.; Gernaey, K.V. A flexible well-mixed milliliter-scale reactor with high oxygen transfer rate for microbial cultivations. Chem. Eng. J. 2016, 303, 655–666. [Google Scholar] [CrossRef]
- Girard, P.; Jordan, M.; Tsao, M.; Wurm, F.M.M. Small-scale bioreactor system for process development and optimization. Biochem. Eng. J. 2001, 7, 117–119. [Google Scholar] [CrossRef]
- Isett, K.; George, H.; Herber, W.; Amanullah, A. Twenty-four-well plate miniature bioreactor high-throughput system: Assessment for microbial cultivations. Biotechnol. Bioeng. 2007, 98, 1017–1028. [Google Scholar] [CrossRef]
- Kensy, F.; Zang, E.; Faulhammer, C.; Tan, R.-K.; Büchs, J. Validation of a high-throughput fermentation system based on online monitoring of biomass and fluorescence in continuously shaken microtiter plates. Microb. Cell Fact. 2009, 8, 31. [Google Scholar] [CrossRef]
- Lladó Maldonado, S.; Panjan, P.; Sun, S.; Rasch, D.; Sesay, A.M.; Mayr, T.; Krull, R. A fully online sensor-equipped, disposable multiphase microbioreactor as a screening platform for biotechnological applications. Biotechnol. Bioeng. 2019, 116, 65–75. [Google Scholar] [CrossRef]
- Kostov, Y.; Harms, P.; Randers-Eichhorn, L.; Rao, G. Low-cost microbioreactor for high-throughput bioprocessing. Biotechnol. Bioeng. 2001, 72, 346–352. [Google Scholar] [CrossRef]
- Doig, S.D.; Diep, A.; Baganz, F. Characterisation of a novel miniaturised bubble column bioreactor for high throughput cell cultivation. Biochem. Eng. J. 2005, 23, 97–105. [Google Scholar] [CrossRef]
- Tsai, C.-H.; Wu, X.; Kuan, D.-H.; Zimmermann, S.; Zengerle, R.; Koltay, P. Digital hydraulic drive for microfluidics and miniaturized cell culture devices based on shape memory alloy actuators. J. Micromech. Microeng. 2018, 28, 084001. [Google Scholar] [CrossRef]
- Lee, K.S.; Boccazzi, P.; Sinskey, A.J.; Ram, R.J. Microfluidic chemostat and turbidostat with flow rate, oxygen, and temperature control for dynamic continuous culture. Lab Chip 2011, 11, 1730–1739. [Google Scholar] [CrossRef] [PubMed]
- Hemmerich, J.; Noack, S.; Wiechert, W.; Oldiges, M. Microbioreactor systems for accelerated bioprocess development. Biotechnol. J. 2018, 13, 1700141. [Google Scholar] [CrossRef] [PubMed]
- Ladner, T.; Grünberger, A.; Probst, C.; Kohlheyer, D.; Büchs, J.; Delvigne, F. Application of mini- and micro-bioreactors for microbial bioprocesses. In Current Developments in Biotechnology and Bioengineering: Bioprocesses, Bioreactors and Controls; Elsevier: Amsterdam, The Netherlands, 2017; pp. 433–461. ISBN 9780444636744. [Google Scholar]
- US Department of Health and Human Services; Food and Drug Administration; Center for Drug Evaluation and Research; Center for Veterinary Medicine; Office of Regulatory Affairs. Guidance for Industry PAT: A Framework for Innovative Pharmaceutical Development, Manufacuring, and Quality Assurance; Food and Drug Administration: Rockville, MD, USA, 2004.
- Gernaey, K.V.; Baganz, F.; Franco-Lara, E.; Kensy, F.; Krühne, U.; Luebberstedt, M.; Marx, U.; Palmqvist, E.; Schmid, A.; Schubert, F.; et al. Monitoring and control of microbioreactors: An expert opinion on development needs. Biotechnol. J. 2012, 7, 1308–1314. [Google Scholar] [CrossRef]
- Frey, L.J.; Vorländer, D.; Rasch, D.; Meinen, S.; Müller, B.; Mayr, T.; Dietzel, A.; Grosch, J.H.; Krull, R. Defining mass transfer in a capillary wave micro-bioreactor for dose-response and other cell-based assays. Biochem. Eng. J. 2020, 161, 107667. [Google Scholar] [CrossRef]
- Zanzotto, A.; Szita, N.; Boccazzi, P.; Lessard, P.; Sinskey, A.J.; Jensen, K.F. Membrane-aerated microbioreactor for high-throughput bioprocessing. Biotechnol. Bioeng. 2004, 87, 243–254. [Google Scholar] [CrossRef]
- Rowland-Jones, R.C.; Jaques, C. At-line raman spectroscopy and design of experiments for robust monitoring and control of miniature bioreactor cultures. Biotechnol. Prog. 2019, 35, e2740. [Google Scholar] [CrossRef]
- Goldrick, S.; Umprecht, A.; Tang, A.; Zakrzewski, R.; Cheeks, M.; Turner, R.; Charles, A.; Les, K.; Hulley, M.; Spencer, C.; et al. High-throughput Raman spectroscopy combined with innovate data analysis workflow to enhance biopharmaceutical process development. Processes 2020, 8, 1179. [Google Scholar] [CrossRef]
- Luchterhand, B.; Nolten, J.; Hafizovic, S.; Schlepütz, T.; Wewetzer, S.J.; Pach, E.; Meier, K.; Wandrey, G.; Büchs, J. Newly designed and validated impedance spectroscopy setup in microtiter plates successfully monitors viable biomass online. Biotechnol. J. 2015, 10, 1259–1268. [Google Scholar] [CrossRef]
- Hofmann, M.C.; Funke, M.; Büchs, J.; Mokwa, W.; Schnakenberg, U. Development of a four electrode sensor array for impedance spectroscopy in high content screenings of fermentation processes. Sens. Actuators B Chem. 2010, 147, 93–99. [Google Scholar] [CrossRef]
- Spiller, E.; Frömmichen, T.; Zimmermann, A.; Sippel, A.E.; Urban, G.A. Development of an electronic microtiterplate for high throughput screening (HTS). Proc. IEEE Sens. 2003, 2, 1041–1044. [Google Scholar] [CrossRef]
- Zanzotto, A.; Boccazzi, P.; Gorret, N.; Van Dyk, T.K.; Sinskey, A.J.; Jensen, K.F. In situ measurement of bioluminescence and fluorescence in an integrated microbioreactor. Biotechnol. Bioeng. 2006, 93, 40–47. [Google Scholar] [CrossRef] [PubMed]
- Samorski, M.; Müller-Newen, G.; Büchs, J. Quasi-continuous combined scattered light and fluorescence measurements: A novel measurement technique for shaken microtiter plates. Biotechnol. Bioeng. 2005, 92, 61–68. [Google Scholar] [CrossRef] [PubMed]
- Faassen, S.M.; Hitzmann, B. Fluorescence spectroscopy and chemometric modeling for bioprocess monitoring. Sensors 2015, 15, 10271–10291. [Google Scholar] [CrossRef] [PubMed]
- Rathore, A.S.; Bhushan, N.; Hadpe, S. Chemometrics applications in biotech processes: A review. Biotechnol. Prog. 2011, 27, 307–315. [Google Scholar] [CrossRef]
- Mowbray, M.; Savage, T.; Wu, C.; Song, Z.; Cho, B.A.; Del Rio-Chanona, E.A.; Zhang, D. Machine learning for biochemical engineering: A review. Biochem. Eng. J. 2021, 172, 108054. [Google Scholar] [CrossRef]
- Wold, S.; Esbensen, K.; Geladi, P. Principal component analysis. Chemom. Intell. Lab. Syst. 1987, 2, 37–52. [Google Scholar] [CrossRef]
- Solle, D.; Hitzmann, B.; Herwig, C.; Pereira Remelhe, M.; Ulonska, S.; Wuerth, L.; Prata, A.; Steckenreiter, T. Between the poles of data-driven and mechanistic modeling for process operation. Chem. Ing. Tech. 2017, 89, 542–561. [Google Scholar] [CrossRef]
- Kirdar, A.O.; Chen, G.; Weidner, J.; Rathore, A.S. Application of near-infrared (NIR) spectroscopy for screening of raw materials used in the cell culture medium for the production of a recombinant therapeutic protein. Biotechnol. Prog. 2010, 26, 527–531. [Google Scholar] [CrossRef]
- Rhee, J.I.; Kang, T.-H.; Lee, K.-I.; Sohn, O.-J.; Kim, S.-Y.; Chung, S.-W. Application of principal component analysis and self-organizing map to the analysis of 2D fluorescence spectra and the monitoring of fermentation processes. Biotechnol. Bioprocess Eng. 2006, 11, 432–441. [Google Scholar] [CrossRef]
- Graf, A.; Claßen, J.; Solle, D.; Hitzmann, B.; Rebner, K.; Hoehse, M. A novel LED-based 2D-fluorescence spectroscopy system for in-line monitoring of chinese hamster ovary cell cultivations–Part I. Eng. Life Sci. 2019, 19, 352–362. [Google Scholar] [CrossRef] [PubMed]
- Hans, S.; Ulmer, C.; Narayanan, H.; Brautaset, T.; Krausch, N.; Neubauer, P.; Schäffl, I.; Sokolov, M.; Cruz Bournazou, M.N. Monitoring parallel robotic cultivations with online multivariate analysis. Processes 2020, 8, 582. [Google Scholar] [CrossRef]
- Mora, A.; Zhang, S.S.; Carson, G.; Nabiswa, B.; Hossler, P.; Yoon, S. Sustaining an efficient and effective CHO cell line development platform by incorporation of 24-deep well plate screening and multivariate analysis. Biotechnol. Prog. 2018, 34, 175–186. [Google Scholar] [CrossRef] [PubMed]
- Ladner, T.; Mühlmann, M.; Schulte, A.; Wandrey, G.; Büchs, J. Prediction of Escherichia coli expression performance in microtiter plates by analyzing only the temporal development of scattered light during culture. J. Biol. Eng. 2017, 11, 20. [Google Scholar] [CrossRef] [PubMed]
- Ödman, P.; Johansen, C.L.; Olsson, L.; Gernaey, K.V.; Lantz, A.E. On-line estimation of biomass, glucose and ethanol in Saccharomyces cerevisiae cultivations using in-situ multi-wavelength fluorescence and software sensors. J. Biotechnol. 2009, 144, 102–112. [Google Scholar] [CrossRef]
- Wold, S.; Sjöström, M.; Eriksson, L. PLS-regression: A basic tool of chemometrics. Chemom. Intell. Lab. Syst. 2001, 58, 109–130. [Google Scholar] [CrossRef]
- Lourenço, N.D.; Lopes, J.A.; Almeida, C.F.; Sarraguça, M.C.; Pinheiro, H.M. Bioreactor monitoring with spectroscopy and chemometrics: A review. Anal. Bioanal. Chem. 2012, 404, 1211–1237. [Google Scholar] [CrossRef]
- Claßen, J.; Aupert, F.; Reardon, K.F.; Solle, D.; Scheper, T. Spectroscopic sensors for in-line bioprocess monitoring in research and pharmaceutical industrial application. Anal. Bioanal. Chem. 2017, 409, 651–666. [Google Scholar] [CrossRef]
- Teixeira, A.P.; Duarte, T.M.; Oliveira, R.; Carrondo, M.J.T.; Alves, P.M. High-throughput analysis of animal cell cultures using two-dimensional fluorometry. J. Biotechnol. 2011, 151, 255–260. [Google Scholar] [CrossRef]
- Kosa, G.; Shapaval, V.; Kohler, A.; Zimmermann, B. FTIR spectroscopy as a unified method for simultaneous analysis of intra- and extracellular metabolites in high-throughput screening of microbial bioprocesses. Microb. Cell Fact. 2017, 16, 195. [Google Scholar] [CrossRef] [Green Version]
- Rowland-Jones, R.C.; Graf, A.; Woodhams, A.; Diaz-Fernandez, P.; Warr, S.; Soeldner, R.; Finka, G.; Hoehse, M. Spectroscopy integration to miniature bioreactors and large scale production bioreactors–Increasing current capabilities and model transfer. Biotechnol. Prog. 2020, 37, e3074. [Google Scholar] [CrossRef] [PubMed]
- Sawatzki, A.; Hans, S.; Narayanan, H.; Haby, B.; Krausch, N.; Sokolov, M.; Glauche, F.; Riedel, S.L.; Neubauer, P.; Bournazou, M.N.C. Accelerated bioprocess development of endopolygalacturonase-production with Saccharomyces cerevisiae using multivariate prediction in a 48 mini-bioreactor automated platform. Bioengineering 2018, 5, 101. [Google Scholar] [CrossRef] [PubMed]
- Ladner, T.; Beckers, M.; Hitzmann, B.; Büchs, J. Parallel online multi-wavelength (2D) fluorescence spectroscopy in each well of a continuously shaken microtiter plate. Biotechnol. J. 2016, 11, 1605–1616. [Google Scholar] [CrossRef] [PubMed]
- Paquet-Durand, O.; Ladner, T.; Büchs, J.; Hitzmann, B. Calibration of a chemometric model by using a mathematical process model instead of offline measurements in case of a H. polymorpha cultivation. Chemom. Intell. Lab. Syst. 2017, 171, 74–79. [Google Scholar] [CrossRef]
- Geinitz, B.; Rehmann, L.; Büchs, J.; Regestein, L. Noninvasive tool for optical online monitoring of individual biomass concentrations in a defined coculture. Biotechnol. Bioeng. 2020, 117, 999–1011. [Google Scholar] [CrossRef]
- Jeude, M.; Dittrich, B.; Niederschulte, H.; Anderlei, T.; Knocke, C.; Klee, D.; Büchs, J. Fed-batch mode in shake flasks by slow-release technique. Biotechnol. Bioeng. 2006, 95, 433–445. [Google Scholar] [CrossRef]
- Flitsch, D.; Krabbe, S.; Ladner, T.; Beckers, M.; Schilling, J.; Mahr, S.; Conrath, U.; Schomburg, W.K.; Büchs, J. Respiration activity monitoring system for any individual well of a 48-well microtiter plate. J. Biol. Eng. 2016, 10, 14. [Google Scholar] [CrossRef]
- Kucheryavskiy, S. Multivariate Data Analysis Toolbox for MATLAB (V.0.1.6). Available online: https://github.com/svkucheryavski/mdatoolsm (accessed on 14 March 2020).
- Jong, S. SIMPLS: An alternative approach to partial least squares regression. Chemom. Intell. Lab. Syst. 1993, 18, 251–263. [Google Scholar] [CrossRef]
- Lakowicz, J.R. Principles of Fluorescence Spectroscopy, 3rd ed.; Springer Science & Business Media: Baltimore, MD, USA, 2006; ISBN 978-0-387-31278-1. [Google Scholar] [CrossRef]
- Cormack, B.; Valdivia, R.; Falkow, S. FACS-optimized mutants of the green fluorescent protein (GFP). Gene 1996, 173, 33–38. [Google Scholar] [CrossRef]
- Tsien, R.Y. The green fluorescent protein. Annu. Rev. Biochem. 1998, 67, 509–544. [Google Scholar] [CrossRef]
- Ghisla, S.; Massey, V.; Lhoste, J.M.; Mayhew, S.G. Fluorescence and optical characteristics of reduced flavines and flavoproteins. Biochemistry 1974, 13, 589–597. [Google Scholar] [CrossRef] [PubMed]
- Heim, R.; Prasher, D.C.; Tsien, R.Y. Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc. Natl. Acad. Sci. USA 1994, 91, 12501–12504. [Google Scholar] [CrossRef] [PubMed]
- Ramanujam, N. Fluorescence spectroscopy of neoplastic and non-neoplastic tissues. Neoplasia 2000, 2, 89–117. [Google Scholar] [CrossRef] [PubMed]
- Beckman Coulter GmbH Round Well Plate-Technical Data Sheet. Available online: https://www.m2p-labs.com/bioreactors/microtiter-plates/round-well-plate/ (accessed on 3 August 2022).
- Kottmeier, K.; Weber, J.; Müller, C.; Bley, T.; Büchs, J. Asymmetric division of Hansenula polymorpha reflected by a drop of light scatter intensity measured in batch microtiter plate cultivations at phosphate limitation. Biotechnol. Bioeng. 2009, 104, 554–561. [Google Scholar] [CrossRef]
- Kunze, M.; Roth, S.; Gartz, E.; Büchs, J. Pitfalls in optical on-line monitoring for high-throughput screening of microbial systems. Microb. Cell Fact. 2014, 13, 53–73. [Google Scholar] [CrossRef]
- Gellissen, G.; Hollenberg, C.P. Application of yeasts in gene expression studies: A comparison of Saccharomyces cerevisiae, Hansenula polymorpha and Kluyveromyces lactis-A review. Gene 1997, 190, 87–97. [Google Scholar] [CrossRef]
- Eggeling, L.; Sahm, H. Derepression and partial insensitivity to carbon catabolite repression of the methanol dissimilating enzymes in Hansenula polymorpha. Eur. J. Appl. Microbiol. Biotechnol. 1978, 5, 197–202. [Google Scholar] [CrossRef]
- Hartner, F.S.; Glieder, A. Regulation of methanol utilisation pathway genes in yeasts. Microb. Cell Fact. 2006, 5, 39. [Google Scholar] [CrossRef]
- Bayer, B.; Sissolak, B.; Duerkop, M.; von Stosch, M.; Striedner, G. The shortcomings of accurate rate estimations in cultivation processes and a solution for precise and robust process modeling. Bioprocess Biosyst. Eng. 2020, 43, 169–178. [Google Scholar] [CrossRef] [Green Version]
- Bratbak, G.; Dundas, I. Bacterial dry matter content and biomass estimations. Appl. Environ. Microbiol. 1984, 48, 755–757. [Google Scholar] [CrossRef]
- Geladi, P.; Kowalski, B.R. Partial least-squares regression: A tutorial. Anal. Chim. Acta 1986, 185, 1–17. [Google Scholar] [CrossRef]
- Abdi, H. Partial least squares regression and projection on latent structure regression (PLS Regression). Wiley Interdiscip. Rev. Comput. Stat. 2010, 2, 97–106. [Google Scholar] [CrossRef]
- Brunner, V.; Siegl, M.; Geier, D.; Becker, T. Challenges in the development of soft sensors for bioprocesses: A critical review. Front. Bioeng. Biotechnol. 2021, 9, 722202. [Google Scholar] [CrossRef] [PubMed]
- Molinaro, A.M.; Simon, R.; Pfeiffer, R.M. Prediction error estimation: A comparison of resampling methods. Bioinformatics 2005, 21, 3301–3307. [Google Scholar] [CrossRef]
- Teixeira, A.P.; Portugal, C.A.M.; Carinhas, N.; Dias, J.M.L.; Crespo, J.P.; Alves, P.M.; Carrondo, M.J.T.; Oliveira, R. In situ 2D fluorometry and chemometric monitoring of mammalian cell cultures. Biotechnol. Bioeng. 2009, 102, 1098–1106. [Google Scholar] [CrossRef] [PubMed]
- Skibsted, E.; Lindemann, C.; Roca, C.; Olsson, L. On-line bioprocess monitoring with a multi-wavelength fluorescence sensor using multivariate calibration. J. Biotechnol. 2001, 88, 47–57. [Google Scholar] [CrossRef]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Berg, C.; Ihling, N.; Finger, M.; Paquet-Durand, O.; Hitzmann, B.; Büchs, J. Online 2D Fluorescence Monitoring in Microtiter Plates Allows Prediction of Cultivation Parameters and Considerable Reduction in Sampling Efforts for Parallel Cultivations of Hansenula polymorpha. Bioengineering 2022, 9, 438. https://doi.org/10.3390/bioengineering9090438
Berg C, Ihling N, Finger M, Paquet-Durand O, Hitzmann B, Büchs J. Online 2D Fluorescence Monitoring in Microtiter Plates Allows Prediction of Cultivation Parameters and Considerable Reduction in Sampling Efforts for Parallel Cultivations of Hansenula polymorpha. Bioengineering. 2022; 9(9):438. https://doi.org/10.3390/bioengineering9090438
Chicago/Turabian StyleBerg, Christoph, Nina Ihling, Maurice Finger, Olivier Paquet-Durand, Bernd Hitzmann, and Jochen Büchs. 2022. "Online 2D Fluorescence Monitoring in Microtiter Plates Allows Prediction of Cultivation Parameters and Considerable Reduction in Sampling Efforts for Parallel Cultivations of Hansenula polymorpha" Bioengineering 9, no. 9: 438. https://doi.org/10.3390/bioengineering9090438