The goal of this literature review is to find the most used commercial bioreactors in medical laboratories, find the most used functions of commercial laboratory bioreactors in the medical industry, identify the differences between our current open source bioreactor and these commercial bioreactors, and identify what changes should be made to our current design to ensure a fair amount of functionality while maintaining open source accessibility.
Citation List For Papers Detailing Medical Laboratory Use of Bioreactors[edit | edit source]
- Tikhomirova, T. S., M. S. Taraskevich, and O. V. Ponomarenko. "The Role of Laboratory-Scale Bioreactors at the Semi-Continuous and Continuous Microbiological and Biotechnological Processes." Applied Microbiology and Biotechnology 102, no. 17 (September 1, 2018): 7293–7308. https://doi.org/10.1007/s00253-018-9194-z.
- Great review that following 13 references are pulled from
- Contains a table of used medical bioreactors, with one citation each
- Contains a few uses of bioreactor in practice: Penicillin (Penicillium chrysogenum), C. cylindracea ATCC 14830 is the producer of extracellular lipases, CHO cell lines are the producer of monoclonal antibodies, site-specific antibody-drug conjugates, or fusion proteins.
- Guilherme, Ederson Paulo Xavier, Jocilane Pereira de Oliveira, Lorendane Millena de Carvalho, Igor Viana Brandi, Sérgio Henrique Sousa Santos, Gleidson Giordano Pinto de Carvalho, Junio Cota, and Bruna Mara Aparecida de Carvalho. "Synthesis of Supermacroporous Cryogel for Bioreactors Continuous Starch Hydrolysis." ELECTROPHORESIS 38, no. 22–23 (2017): 2940–46. https://doi.org/10.1002/elps.201700208.
- In this work, a new bioreactor was synthesized using cryogel monoliths as support material for the immobilization of the alpha amylase from A. oryzae. The bioreactor continuously produced maltose from potato starch.
- Frensing, Timo, Frank Stefan Heldt, Antje Pflugmacher, Ilona Behrendt, Ingo Jordan, Dietrich Flockerzi, Yvonne Genzel, and Udo Reichl. "Continuous Influenza Virus Production in Cell Culture Shows a Periodic Accumulation of Defective Interfering Particles." PLOS ONE 8, no. 9 (September 5, 2013): e72288. https://doi.org/10.1371/journal.pone.0072288.
- A two-stage bioreactor setup was designed in which cells were cultivated in a first stirred tank reactor where an almost constant cell concentration was maintained. Cells were then constantly fed to a second bioreactor where virus infection and replication took place. Using this two-stage reactor system, it was possible to continuously produce influenza viruses.
- . Even with very low amounts of DIPs in the seed virus and very low rates for de novo DIP generation, defective viruses rapidly accumulate and, therefore, represent a serious challenge for continuous vaccine production. Yet, the continuous replication of influenza virus using a two-stage bioreactor setup is a novel tool to study aspects of viral evolution and the impact of DIPs
- Two small scale stirred tank bioreactors (1 L working volume Biostat B plus, Sartorius) were used.
- The first lab-scale bioreactor was inoculated with AGE1.CR cells and cultivations were carried out at 37uC, pH 7.2 and a stirring speed of 120 rpm with a working volume of 1 L. Aeration was controlled to 40% DO by pulsed aeration with pure oxygen through a microsparger (whole methods section details a medical use of bioreactor and specifics)
- Tapia, Felipe, Ingo Jordan, Yvonne Genzel, and Udo Reichl. "Efficient and Stable Production of Modified Vaccinia Ankara Virus in Two-Stage Semi-Continuous and in Continuous Stirred Tank Cultivation Systems." PLOS ONE 12, no. 8 (August 24, 2017): e0182553. https://doi.org/10.1371/journal.pone.0182553.
- In this work, continuous production of Modified Vaccinia Ankara (MVA) virus was investigated.
- The process was automated in a two-stage continuous system comprising two connected 1 L stirred tank bioreactors.
- A bioreactor system consisting of two 1 L stirred tank bioreactors (Biostat B Plus, Sartorius) was established (similar to Frensing et al. 2013, Fig 1B). The first bioreactor (Cell Bioreactor, CB) was inoculated with AGE1.CR.pIX cells at 1×106 cells/mL and operated at 37˚C, 120 rpm with Rushton impellers, 40% oxygen saturation and 850 mL working volume (wv). Oxygen saturation was controlled using pulsed aeration with pure oxygen. The pH value was not controlled and only monitored during the batch phase to avoid values below 6.9 (Methods Detailed again)
- 37˚C
- oxygen concentration was at 40–50% saturation
- MVA-CR19 virus was added to VB at an moi of 0.05
- The peristaltic pumps used were Ismatec Reglo-Digital MS2/8-160 (Pump 1 and 2, Fig 1B; Cole-Parmer GmbH, Germany), and Watson Marlow 101U/R (Pump 3, Fig 1B; Waston-Marlow Fluid Technology Group, UK).
- Wu, Hsuan-Chen, Yu-Chen Hu, and William E. Bentley. "Tubular Bioreactor for Probing Baculovirus Infection and Protein Production." In Baculovirus and Insect Cell Expression Protocols, edited by David W. Murhammer, 461–67. Methods in Molecular Biology. New York, NY: Springer, 2016. https://doi.org/10.1007/978-1-4939-3043-2_23.
- This chapter describes several alternative bioreactor systems for baculovirus infection. We provide an example alternative system that holds promise to avoid asynchronous distributions in infection time. Namely, we describe a two-stage reactor system consisting of an upstream continuous stirred tank reactor and a downstream tubular reactor with segmented plug fl ow for probing baculovirus infection and production.
- House made two stage continuous bioreactor: Culture room controlled at 27–28 °C., A glass-blown jacketed spinner fl ask (working volume = 200 mL) made in house., One-liter medium bottles with multi-port cap assemblies (Bellco Biotechnology) to serve as medium reservoir and waste jar., Stirrer plate., Water bath., Needles (22-gauge) and syringe (1 mL) for sampling., Rubber stopper (No. 10)., Stirrer assembly (Bellco Biotechnology)., Masterfl ex precision L/S peristaltic pumps (Cole Palmer)., Microprocessor-controlled peristaltic multichannel pump (Watson Marlow, model 505Du) and a 4-channel pump head (model 205BA)., Watson-Marlow silicone tubing (ID = 1.14 mm) and MasterFlex no. 14 tubing (length = 7.6 m, ID = 1.6 mm).
- O'Mara, P., A. Farrell, J. Bones, and K. Twomey. "Staying Alive! Sensors Used for Monitoring Cell Health in Bioreactors." Talanta 176 (January 1, 2018): 130–39. https://doi.org/10.1016/j.talanta.2017.07.088.
- A number of advances in sensor technology have been achieved in recent years, a few of these advances and future research will also be discussed in this review.
- a large number of recombinant monoclonal products have been brought to market such as Rituxan, Herceptin and Remicade
- pH is one of the 3 crucial parameters that must be closely monitored and controlled to prevent cell apoptosis from occurring. pH for optimal cell growth is usually about 7.6 for most animal cells but fluctuations do occur during the cell cycle with pH's reaching levels of 7.0 in some cases
- Cell culture media normally contains buffer agents and sodium bicarbonates to keep pH within optimal working parameters.
- Combined with CO2 sparging to reduce pH and base to increase it. It is also noted that optimal pH changes over the course of a bioprocess.
- even a small change of 0.1 pH units from the optimum can have a large impact on cell viability and concentration.
- Considering that tests have shown that a change of as little as 0.15 pH units can result in a reduction of protein expression levels, so it is highly important that these probes are highly accurate [34].
- Dissolved oxygen (dO) is another key parameter that must be closely monitored and optimised in cell production in a bioreactor.
- Temperature monitoring is crucial to ensure optimal cell viability and product yield during bioprocessing. For mammalian cells the optimal temperature for production has been understood for a number of years to be around 37 °C
- d. However recent studies have shown that lower temperature in the range of 30–35 °C could yield high productions of some protein types.
- over 37 °c caused a great loss in cell viability and cellular production
- Therefore, it is clear that temperature sensors must operate accurately in a range of 30–40 °c as the process temperature will change over time. Due to this small margin these sensors must be pinpoint accurate to avoid loss in cell viability
- Fazenda, Mariana L., and Linda M. Harvey and Brian McNeil. "Effects of Dissolved Oxygen on Fungal Morphology and Process Rheology during Fed-Batch Processing of Ganoderma Lucidum" 20, no. 4 (April 28, 2010): 844–51. https://www.jmb.or.kr/journal/view.html?spage=844&volume=20&number=4.
- The composition of the medium used in all stages was (g/l): glucose 35, yeast extract 5, peptone 5, KH2PO4 1, and MgSO4·7H2O 0.5. All fermentations were carried out in fed-batch mode with pulse-feeding of glucose, yeast extract, and peptone every 24 h, or whenever the glucose concentration fell to 5-10 g/l. The reactor used was a 15-l (total volume) stainless steel bioreactor (BIOSTAT C.-DCU; B. Braun Biotech International, Switzerland). The pH was kept at 4.0 by automatic addition of titrants (1 M NaOH and 1 M H2SO4). The temperature was kept at 30o C throughout the runs and the agitation rate was set at 300 rpm. The culture conditions chosen were based on previous work [7]. 30o C and 300 rpm Uncontrolled and controlled dissolved oxygen (DO) fed-batch cultures were performed. In the DOcontrolled process, the DO set-point was at 20% air saturation using a cascade control of agitation, airflow rate, and oxygen enrichment. The maximum airflow rate used was at 2.0 volume of air per volume of culture per minute (vvm) in both cultures. Real-time values of pH, dissolved oxygen, agitation speed, temperature, airflow rate, and oxygen percentage during fermentations were recorded automatically by the bioreactor software, MFCS DA (Sartorius, U.K.)
- Tescione, Lia, James Lambropoulos, Madhava Ram Paranandi, Helena Makagiansar, and Thomas Ryll. "Application of Bioreactor Design Principles and Multivariate Analysis for Development of Cell Culture Scale down Models." Biotechnology and Bioengineering 112, no. 1 (2015): 84–97. https://doi.org/10.1002/bit.25330.
- The production process was operated in a 3-L, stirred tank bioreactor (Applikon Biotechnology, Schiedam, Netherlands) as a fed-batch culture in serum-free medium.
- Temperature, pH, and dissolved oxygen were maintained using TruViu RDPD utility tower, TruLogic controllers, and TruBio software (Finesse Solutions, Santa Clara, CA). Temperature was controlled at 35�C. pH was controlled at 7.2 (�0.1 deadband) through addition of CO2 or 1.0 molal sodium carbonate. Dissolved oxygen was controlled by delivery of air and/or oxygen through a drilled hole sparger or through sintered spargers with a 15 or 50 mm pore size. Antifoam was added as 10 ppm bolus shots to reduce foam accumulation and avoid exhaust filter wetting
- Theodore, Christine M., Steven T. Loveridge, Mitchell S. Crews, Nicholas Lorig-Roach, and Phillip Crews. "Design and Implementation of an Affordable Laboratory-Scale Bioreactor for the Production of Microbial Natural Products." Engineering Reports 1, no. 4 (2019): e12059. https://doi.org/10.1002/eng2.12059.
- To address these challenges a cost effective, laboratory scale bioreactor was designed and implemented. The constructed bioreactor addresses common problems that small or teaching-focused laboratories face when attempting scale up cultures.
- Xu, Ping, Colleen Clark, Todd Ryder, Colleen Sparks, Jiping Zhou, Michelle Wang, Reb Russell, and Charo Scott. "Characterization of TAP Ambr 250 Disposable Bioreactors, as a Reliable Scale-down Model for Biologics Process Development." Biotechnology Progress 33, no. 2 (2017): 478–89. https://doi.org/10.1002/btpr.2417.
- The three cell lines were thawed and expanded in proprietary chemically defined medium at 378C with 5% CO2 overlay prior to inoculation in Ambr 250, 5-L, and 250-L bioreactors.
- About 5-L bench-top bioreactors (Sartorius Stedim Biotech) were used in upstream process development. The processes were scaled up in 250-L single-use bioreactors
- Kumar, Vinod, Vasudha Bansal, Aravind Madhavan, Manoj Kumar, Raveendran Sindhu, Mukesh Kumar Awasthi, Parameswaran Binod, and Saurabh Saran. "Active Pharmaceutical Ingredient (API) Chemicals: A Critical Review of Current Biotechnological Approaches." Bioengineered 13, no. 2 (February 1, 2022): 4309–27. https://doi.org/10.1080/21655979.2022.2031412.
- The aim of this article was to generate a framework of bio-based economy by an effective utilization of biomass from the perspectives of agriculture for developing potential end biobased products (e.g. pharmaceuticals, active pharmaceutical ingredients). Our discussion is also extended to the conservatory ways of bioenergy along with development of bio-based products and biofuels. This review article further showcased the fundamental principles for producing these by-products.
- table 1 has many chemicals and their production strategy
- also some important APIs
- Xu, Sen, Linda Hoshan, Rubin Jiang, Balrina Gupta, Eric Brodean, Kristin O'Neill, T. Craig Seamans, John Bowers, and Hao Chen. "A Practical Approach in Bioreactor Scale-up and Process Transfer Using a Combination of Constant P/V and Vvm as the Criterion." Biotechnology Progress 33, no. 4 (July 2017): 1146–59. https://doi.org/10.1002/btpr.2489.
- The inoculum trains started from vial thaws and were expanded in shake flasks followed by rocking wave bioreactors (WAVE Bio�reactorTM 20/50, GE Healthcare, Uppsala, Sweden, or BIO�STATVR RM 20/50, Sartorius Stedim, G€ottingen, Germany) with increasing volumes.
- Glass bioreactors (3 L, Sartorius Stedim, G€ottingen, Ger�many) with a single marine impeller were used with a drilled hole sparger (DHS) or a frit sparger. SS bioreactors at 500 L (Sites A and C) or 2,000 L (Sites B and C) with a single DHS were used. SUBs at 200 L and 2,000 L (Xcellerex XDR200 and XDR2000, GE Healthcare, Marlborough, MA) and 500 L (Hyclone 500 L S.U.B., Thermo Fisher Scientific, Logan, UT) (Sites A and D) with a dual sparger system (Frit sparger for O2 and CO2, DHS for air or N2) were used. The tank geometry, sparger design, and impeller configuration varied from one to another (Table 1)
- Zheng, Ming-Min, Fei-Fei Chen, Hao Li, Chun-Xiu Li, and Jian-He Xu. "Continuous Production of Ursodeoxycholic Acid by Using Two Cascade Reactors with Co-Immobilized Enzymes." Chembiochem: A European Journal of Chemical Biology 19, no. 4 (February 16, 2018): 347–53. https://doi.org/10.1002/cbic.201700415.
- Ursodeoxycholic acid (UDCA) is an effective drug for the treat�ment of hepatitis. In this study, 7a-hydroxysteroid dehydrogen�ase (7a-HSDH) and lactate dehydrogenase (LDH), as well as 7b-hydroxysteroid dehydrogenase (7b-HSDH) and glucose de�hydrogenase (GDH), were co-immobilized onto an epoxy-func�tionalized resin (ES-103) to catalyze the synthesis of UDCA from chenodeoxycholic acid (CDCA).
- two serial packed-bed reactors.
- Middleton, Brennen. "Design of an Undergraduate Laboratory Experiment Utilizing a Stirred-Tank, Jacketed Bioreactor," n.d., 96.
- For the proposed laboratory experiment, a BIOSTAT© M reactor, a jacketed, continuous-stirred batch reactor, with a 1.5 L working volume manufactured by Sartorius will be utilized
List of Top Reusable Stir Tank Bioreactors[edit | edit source]
When searched on google scholar with: "(bioreactor name)" as of May 20th
minifors: 654 results
multifors: 571 results
labfors: 1010 results
Biostat B: 2100 results
Biostat B plus: 1090 results
Biostat C:945 results
Bioflo 110:1960 results
Bioflo III: 1040 results
Bioflo 115: 542 results
Bioflo 310: 371 results
Other bioreactors: Biolafitte, RALF
This method does not accurately represent bioreactor use in academic literature and is only an exploratory tool.
Tikhomirova, T. S., M. S. Taraskevich, and O. V. Ponomarenko. "The Role of Laboratory-Scale Bioreactors at the Semi-Continuous and Continuous Microbiological and Biotechnological Processes." Applied Microbiology and Biotechnology 102, no. 17 (September 1, 2018): 7293–7308. https://doi.org/10.1007/s00253-018-9194-z.
| Denomination | Manufacturer | Working volume, l | Cultivation mode | Stirring mode | Reference |
|---|---|---|---|---|---|
| BIOSTAT®Cplus | Sartorius staedium biotech | 5, 10, 15, 20, 30 | Batch | Rushton turbine, 6 blades | (Fazenda et al. 2010) |
| BIOSTAT® B | Sartorius staedium biotech | 1, 2, 5, 10 | Batch, semi-batch | Rushton turbine, 3 blades | (Orr et al. 2009; Zheng et al. 2017) |
| BIOSTAT® Bplus | Sartorius staedium biotech | 1, 2, 5, 10 | Batch, semi-batch | Rushton turbine, 2 blades | (Frensing et al. 2013; Timoumi et al. 2017b; Tapia et al. 2017; Timoumi et al. 2017a) |
| Braun airlift fermentor (BIOSTAT® B-DCU II) | Sartorius staedium biotech | 2, 5 | Batch, semi-batch | Rushton turbine, Airlift | (Orr et al. 2009; Cobas et al. 2016) |
| New Brunswick FS300 baffled fermenter® | New Brunswick Scientific | 3.5, 5, 7.5, 14, 20 | Batch, semi-batch | Rushton turbine, 6 blades | (Galvagno et al. 2011) |
| BioFlo 3000 | New Brunswick Scientific | 1.6, 3.3, 6.6 | Batch, semi-batch | Rushton turbine, 6 blades | (Nitsche et al. 2012) |
| New Brunswick BioFlo 110 Fermentor | New Brunswick Scientific | 1.3, 3, 7.5, 14 | Batch, semi-batch | Rushton turbine, 2 blades | (Rioseras et al. 2014) |
| Multifors | INFORS AG | 0.25, 0.5, 0.75, 1 | Batch | Rushton turbine, 2 blades | (Jost et al. 2015) |
| Ralf Plus-System | Bioengineering inc. | 2, 3.7, 5, 6.7 | Batch, semi-batch | Rushton turbine, 6 blades | (Bellou et al. 2014) |
| BIOTECH-3BG | Shanghai BaoXing Bio-Engineering Equipment Co., Ltd | 3, 5, 7 | Batch, semi-batch | Rushton turbine, 6 blades | (Chen et al. 2017a) |
| Continuous stirred tank reactors (CSTR) | Nano-Mag Technologies Pvt. Ltd | 1, 2, 5, 10, 20, 25 | Continuous | Rushton turbine, 6 blades | (Zhao et al. 2017) |
| Ambr®250 | Sartorius staedium biotech | 0.25 | Continuous (cascade-mode) | Rushton turbine, 2 blades | (Moore et al. 2017) |
Specifications of Top Medical Bioreactors[edit | edit source]
- Biostat B
- Sensors
- Oxygen (Polarographic or optical)
- PH (Combined measuring electrode), 0.01pH accuracy reusable, 0.1 single use
- Temperature(Pt100)
- Foam control (Electrical conductive sensor)
- Level (Electrical conductive sensor)
- Turbidity (1-channel NIR absorption sensor)
- Redox (Combined measurement with pH sensor | –1,000– 1,000 mV | 1 mV)
- Balance substrate
- Gravimetric Flow Controller
- Balance culture vessel
- RM load cells
- External signal input
- Pump rates: see spec sheet, lowest is 0-0.1ml/min, fastest is 4.25-127.5ml/min
- Flow meters
- Mass Flow Controllers
- Temperature control - electrical heating blanket with cooling finger
- 8-60 Celsius
- 50 Celsius for single use plastic
- Sensors
- Biostat B-dcu
- sensors
- same as biostat B, plus advanced sensors
- pressure
- glucose enzymatic sensor
- lactate enzymatic sensor
- viable biomass capacitance sensor
- O2 offgas, zirconium dioxide
- CO2 offgas, infrared
- Pump rates: see spec sheet
- sensors
- minifors
- sensors
- Oxygen
- PH
- Temperature
- Foam (conductive with dosing needle)
- Turbidity ((Single channel light absorption) or (non-invasive scattered light measurement))
- exit gas flow CO2 and O2
- redox combined sensor instead of pO2
- pump rates, 0.0012 to 1.12 mL min-1
- sensors
- labfors
- sensors
- Oxygen
- PH
- Temperature
- CO2
- pressure
- turbidity
- antifoam
- level
- conductivity/permitivity
- redox
- exit gas analysis
- pump rates, 0.0012 to 1.12 mL min-1
- sensors
- Bioflo 110
- Sensors
- Oxygen
- PH
- Temperature
- foam
- level
- rotameter
- optional exhaust condenser
- pump rates: max 23.5 ml per min, min 0.25ml per min (may be lower if pump has variable speed)
- Sensors
- RALF
- Oxygen
- PH
- Temperature
- M
- Sensors
- Oxygen (1-50 mg/dL)
- PH (0-14)
- Temperature (0-50)
- IR thermometer
- optical density
- Sensors
Most Used Functions of Top Bioreactors[edit | edit source]
- Other than the basic PH, Oxygen, Temperature control, they also control feed substance like glucose, and cell culture density. All of these are already included and accounted for, except for possibly glucose. CO2 is used to control PH with buffers, and N2 gas is used to remove oxygen and CO2 and control PH with buffers. Buffer substances keep a tight PH spread. There is not much our current bioreactor design lacks.
Resulting Possible Changes[edit | edit source]
- Sterilization Needs
- Autoclavable?
- Additional Sensors
- Foam/level
- Glucose
- Accuracy of control
- sensor accuracy
- sensor latency
- pump flow rates
- sparger design
- Single Use Targeting or Multi Use?
- Plastics vs Glass vs Steel
- Software Control Must Ensure Accuracy
Laboratory Applications for Bioreactor Testing[edit | edit source]
- Cultivation of Cell Cultures - CHO cell lines
- Protein Production and Yield testing - E.Coli or other
- Phototropic Organism Cultivation (optional)
- Generic drug production
Other possible drugs we could cultivate:
Ustekinumab - Sp2/0 murine myeloma
Infliximab - mouse myeloma cells (SP2/0 cells)
