Oxygen Transfer Characteristics of Miniaturized Bioreactor Systems

Since their introduction in 2001 miniaturized bioreactor systems have made great advances in function and performance. In this article the dissolved oxygen (DO) transfer performance of submilliliter microbioreactors, and 1–10 mL minibioreactors was examined. Microbioreactors have reached kLa values of 460 h−1, and are offering instrumentation and some functionality comparable to production systems, but at high throughput screening volumes. Minibioreactors, aside from one 1,440 h−1 kLa system, have not offered as high rates of DO transfer, but have demonstrated superior integration with automated fluid handling systems. Microbioreactors have been typically limited to studies with E. coli, while minibioreactors have offered greater versatility in this regard. Further, mathematical relationships confirming the applicability of kLa measurements across all scales have been derived, and alternatives to fluorescence lifetime DO sensors have been evaluated. Finally, the influence on reactor performance of oxygen uptake rate (OUR), and the possibility of its real‐time measurement have been explored. Biotechnol. Bioeng. 2013; 110: 1005–1019. © 2012 Wiley Periodicals, Inc.

[1]  B Allen,et al.  Design of a prototype miniature bioreactor for high throughput automated bioprocessing , 2003 .

[2]  E. Krommenhoek Integrated sensor array for on-line monitoring micro bioreactors , 2007 .

[3]  Ashraf Amanullah,et al.  Twenty‐four well plate miniature bioreactor system as a scale‐down model for cell culture process development , 2009, Biotechnology and bioengineering.

[4]  Yinjie J. Tang,et al.  Evaluation of the effects of various culture conditions on Cr(VI) reduction by Shewanella oneidensis MR‐1 in a novel high‐throughput mini‐bioreactor , 2006, Biotechnology and bioengineering.

[5]  O. Wolfbeis Fiber-optic chemical sensors and biosensors. , 2004, Analytical chemistry.

[6]  Xudong Ge,et al.  Comparisons of optical pH and dissolved oxygen sensors with traditional electrochemical probes during mammalian cell culture , 2007, Biotechnology and bioengineering.

[7]  W. Duetz,et al.  Microtiter plates as mini-bioreactors: miniaturization of fermentation methods. , 2007, Trends in microbiology.

[8]  Armin Fiechter,et al.  Advances in Biochemical Engineering , 1971 .

[9]  Rajeev J Ram,et al.  Microfluidic chemostat and turbidostat with flow rate, oxygen, and temperature control for dynamic continuous culture. , 2011, Lab on a chip.

[10]  Frank Kensy,et al.  The baffled microtiter plate: Increased oxygen transfer and improved online monitoring in small scale fermentations , 2009, Biotechnology and bioengineering.

[11]  I. Marison,et al.  A new method for on-line measurement of the volumetric oxygen uptake rate in membrane aerated animal cell cultures. , 2000, Journal of biotechnology.

[12]  G. Rao,et al.  Low-cost microbioreactor for high-throughput bioprocessing. , 2001, Biotechnology and bioengineering.

[13]  A Lübbert,et al.  Bioreactor performance: a more scientific approach for practice. , 2001, Journal of biotechnology.

[14]  Gerardo Perozziello,et al.  Microchemostat-microbial continuous culture in a polymer-based, instrumented microbioreactor. , 2006, Lab on a chip.

[15]  Yordan Kostov,et al.  Design and performance of a 24‐station high throughput microbioreactor , 2006, Biotechnology and bioengineering.

[16]  Christopher M.A. Brett,et al.  Electrochemistry: Principles, Methods, and Applications , 1993 .

[17]  Ingo Klimant,et al.  Modeling of Mixing in 96‐Well Microplates Observed with Fluorescence Indicators , 2002, Biotechnology progress.

[18]  Dirk Weuster-Botz,et al.  New milliliter‐scale stirred tank bioreactors for the cultivation of mycelium forming microorganisms , 2010, Biotechnology and bioengineering.

[19]  G. Dumsday,et al.  The use of oxygen uptake rate to monitor discovery of microbial and enzymatic biocatalysts , 2009, Biotechnology and bioengineering.

[20]  G J Lye,et al.  Scale‐up of Escherichia coli growth and recombinant protein expression conditions from microwell to laboratory and pilot scale based on matched kLa , 2008, Biotechnology and bioengineering.

[21]  James N. Demas,et al.  Determination of oxygen concentrations by luminescence quenching of a polymer-immobilized transition-metal complex , 1987 .

[22]  George T. Tsao,et al.  Dissolved oxygen electrodes , 1979 .

[23]  Annik Nanchen,et al.  Nonlinear Dependency of Intracellular Fluxes on Growth Rate in Miniaturized Continuous Cultures of Escherichia coli , 2006, Applied and Environmental Microbiology.

[24]  Xudong Ge,et al.  Validation of an optical sensor-based high-throughput bioreactor system for mammalian cell culture. , 2006, Journal of biotechnology.

[25]  J. Büchs,et al.  Characterisation of the gas-liquid mass transfer in shaking bioreactors. , 2001, Biochemical engineering journal.

[26]  A. K. Biń MASS TRANSFER TO THE FREE INTERFACE IN STIRRED VESSELS , 1984 .

[27]  B. Özbek,et al.  The studies on the oxygen mass transfer coefficient in a bioreactor , 2001 .

[28]  J Büchs,et al.  Micro-bioreactors for fed-batch fermentations with integrated online monitoring and microfluidic devices. , 2009, Biosensors & bioelectronics.

[29]  Nicolas Szita,et al.  Membrane‐aerated microbioreactor for high‐throughput bioprocessing , 2004, Biotechnology and bioengineering.

[30]  F. Duschinsky,et al.  Der zeitliche Intensitätsverlauf von intermittierend angeregter Resonanzstrahlung , 1933 .

[31]  Martina Micheletti,et al.  Microscale bioprocess optimisation. , 2006, Current opinion in biotechnology.

[32]  B. Junker Scale-up methodologies for Escherichia coli and yeast fermentation processes. , 2004, Journal of bioscience and bioengineering.

[33]  K. Andersen,et al.  Are growth rates of Escherichia coli in batch cultures limited by respiration? , 1980, Journal of bacteriology.

[34]  Michael C. Flickinger,et al.  encyclopedia of bioprocess technology , 1999 .

[35]  N L Swanson,et al.  Limits of optical transmission measurements with application to particle sizing techniques. , 1999, Applied optics.

[36]  K. Jensen,et al.  In situ measurement of bioluminescence and fluorescence in an integrated microbioreactor. , 2006, Biotechnology and bioengineering.

[37]  O. Wolfbeis,et al.  Fiber-optic chemical sensors and biosensors. , 2008, Analytical chemistry.

[38]  Nicolas Szita,et al.  A well‐mixed, polymer‐based microbioreactor with integrated optical measurements , 2006, Biotechnology and bioengineering.

[39]  Gary J. Lye,et al.  Design and characterisation of a miniature stirred bioreactor system for parallel microbial fermentations , 2008 .

[40]  Christoph Wittmann,et al.  Review: Minibioreactors , 2004, Biotechnology Letters.

[41]  V. Hessel,et al.  Micromixers—a review on passive and active mixing principles , 2005 .

[42]  G. G. Peters,et al.  Mixing in Stokes flow in an annular wedge cavity , 1999 .

[43]  Dirk Weuster-Botz,et al.  Methods and milliliter scale devices for high-throughput bioprocess design , 2005, Bioprocess and biosystems engineering.

[44]  John Alderman,et al.  A low-volume platform for cell-respirometric screening based on quenched-luminescence oxygen sensing. , 2004, Biosensors & bioelectronics.

[45]  Wei-Shou Hu,et al.  High density culture of mammalian cells with dynamic perfusion based on on-line oxygen uptake rate measurements , 2004, Cytotechnology.

[46]  B Sonnleitner,et al.  Growth of Saccharomyces cerevisiae is controlled by its limited respiratory capacity: Formulation and verification of a hypothesis , 1986, Biotechnology and bioengineering.

[47]  B O Palsson,et al.  Growth, Metabolic, and Antibody Production Kinetics of Hybridoma Cell Culture: 2. Effects of Serum Concentration, Dissolved Oxygen Concentration, and Medium pH in a Batch Reactor , 1991, Biotechnology progress.

[48]  Ashraf Amanullah,et al.  Twenty‐four‐well plate miniature bioreactor high‐throughput system: Assessment for microbial cultivations , 2007, Biotechnology and bioengineering.

[49]  A. Nienow Reactor Engineering in Large Scale Animal Cell Culture , 2006, Cytotechnology.

[50]  Harry L. T. Lee,et al.  Polymer waveguide backplanes for optical sensor interfaces in microfluidics. , 2007, Lab on a chip.

[51]  Yordan Kostov,et al.  Bioprocess monitoring. , 2002, Current opinion in biotechnology.

[52]  Gary J. Lye,et al.  Modelling surface aeration rates in shaken microtitre plates using dimensionless groups , 2005 .

[53]  E. Heinzle,et al.  Optical device for parallel online measurement of dissolved oxygen and pH in shake flask cultures , 2010, Bioprocess and biosystems engineering.

[54]  J J Heijnen,et al.  Integrated electrochemical sensor array for on-line monitoring of yeast fermentations. , 2006, Analytical chemistry.

[55]  Mark R. Marten,et al.  Optical analysis of liquid mixing in a minibioreactor , 2006, Biotechnology and bioengineering.

[56]  Elmar Heinzle,et al.  A system of miniaturized stirred bioreactors for parallel continuous cultivation of yeast with online measurement of dissolved oxygen and off‐gas , 2013, Biotechnology and bioengineering.

[57]  Oxygen uptake rate measurements both by the dynamic method and during the process growth of Rhodococcus erythropolis IGTS8: Modelling and difference in results , 2006 .

[58]  Marcel Ottens,et al.  Lab‐scale fermentation tests of microchip with integrated electrochemical sensors for pH, temperature, dissolved oxygen and viable biomass concentration , 2008, Biotechnology and bioengineering.

[59]  D Weuster-Botz,et al.  Development, parallelization, and automation of a gas-inducing milliliter-scale bioreactor for high-throughput bioprocess design (HTBD). , 2005, Biotechnology and bioengineering.

[60]  L. Armstrong,et al.  Theoretical analysis of the phase shift measurement of lifetimes using monochromatic light , 1975 .

[61]  Elmar Heinzle,et al.  On-line oxygen uptake rate and culture viability measurement of animal cell culture using microplates with integrated oxygen sensors , 2004, Biotechnology Letters.

[62]  Elmar Heinzle,et al.  Microplates with integrated oxygen sensing for medium optimization in animal cell culture , 2004, Cytotechnology.

[63]  J. Büchs,et al.  Power consumption in shaking flasks on rotary shaking machines: II. Nondimensional description of specific power consumption and flow regimes in unbaffled flasks at elevated liquid viscosity. , 2000, Biotechnology and bioengineering.

[64]  Krist V. Gernaey,et al.  Development of a single-use microbioreactor for cultivation of microorganisms , 2010 .

[65]  Nicolas Szita,et al.  Development of a multiplexed microbioreactor system for high-throughput bioprocessing. , 2005, Lab on a chip.

[66]  K. Jensen,et al.  Differential Gene Expression Profiles and Real‐Time Measurements of Growth Parameters in Saccharomyces cerevisiae Grown in Microliter‐Scale Bioreactors Equipped with Internal Stirring , 2006, Biotechnology progress.

[67]  J. Nielsen,et al.  Bioreaction Engineering Principles , 1994, Springer US.

[68]  G. Rao,et al.  Comparisons of optically monitored small-scale stirred tank vessels to optically controlled disposable bag bioreactors , 2009, Microbial cell factories.

[69]  Govind Rao,et al.  Phase fluorometric sterilizable optical oxygen sensor , 1994, Biotechnology and bioengineering.

[70]  Jochen Büchs,et al.  Advances in understanding and modeling the gas–liquid mass transfer in shake flasks , 2004 .

[71]  Wilfried Mokwa,et al.  Bioprocess Control in Microscale: Scalable Fermentations in Disposable and User-Friendly Microfluidic Systems , 2010, Microbial cell factories.

[72]  Wouter A. Duetz,et al.  Oxygen transfer by orbital shaking of square vessels and deepwell microtiter plates of various dimensions , 2004 .

[73]  Wilfried Mokwa,et al.  Microfluidic biolector—microfluidic bioprocess control in microtiter plates , 2010, Biotechnology and bioengineering.

[74]  Hua-bei Jiang,et al.  Fluorescence lifetime tomography of turbid media based on an oxygen-sensitive dye. , 2002, Optics express.

[75]  Jonathan I. Betts,et al.  Miniature bioreactors: current practices and future opportunities , 2006, Microbial cell factories.

[76]  J Büchs,et al.  Out-of-phase operating conditions, a hitherto unknown phenomenon in shaking bioreactors. , 2001, Biochemical engineering journal.

[77]  K Konstantinov,et al.  Real-time biomass-concentration monitoring in animal-cell cultures. , 1994, Trends in biotechnology.

[78]  Harry L. T. Lee,et al.  Microbioreactor arrays with integrated mixers and fluid injectors for high-throughput experimentation with pH and dissolved oxygen control. , 2006, Lab on a chip.

[79]  J Büchs,et al.  Power consumption in shaking flasks on rotary shaking machines: I. Power consumption measurement in unbaffled flasks at low liquid viscosity. , 2000, Biotechnology and bioengineering.

[80]  Benjamin A. DeGraff,et al.  Design and Applications of Highly Luminescent Transition Metal Complexes , 1991 .

[81]  Kurt A Brorson,et al.  Advances in clone selection using high‐throughput bioreactors , 2010, Biotechnology progress.

[82]  Klavs F. Jensen,et al.  Gene expression analysis of Escherichia coli grown in miniaturized bioreactor platforms for high-throughput analysis of growth and genomic data , 2005, Applied Microbiology and Biotechnology.

[83]  H. Hang,et al.  Oxygen uptake rate optimization with nitrogen regulation for erythromycin production and scale-up from 50 L to 372 m3 scale. , 2009, Bioresource technology.