1、Bioreactor selectionScale-UpBringing a Process to Scale Each product creates its own unique challenges for process development:o Developing a Biofuels process to the 300,000L scale faces different challenges than developing a Pharmaceutical production process to the 100L-25,000L scale.o Developing a
2、 Pharmaceutical production process in a microbial host faces different challenges than would be faced in a mammalian process.Despite the wide range of differences,most process development efforts require the same basic steps and these steps are a result of shared end goals.What is the Goal?.To run a
3、 multi-million(billion)dollar production facility at full capacity producing end products worth millions(billions)of dollars.How is this Goal Achieved?At the heart of the process is a stable,reliable cell line and culture approach which has been well characterized at a smaller and more manageable sc
4、ale.Create Cell LineSelect in Well-PlatesAdvance to Flasks,Select Further.Expand Cell LineDevelop the ProcessScale the ProcessIdentify ProductR&DLarge Scale ProcessIdentify ProductR&DSelection StepsDevelopment StepsBecause these steps provide very little information to the researcher,“Selection”is p
5、ossible,but true process development is difficult.The data recorded at each step during process development is what allows the researcher to control and therefore scale the process.Information Available at Different Stages.Identify ProductR&DEarly,High-throughput stages provide little information.Re
6、searchers are often limited to Cell Counts and Titers.Process Development stages track and control many parameters:pH,DO,pCO2,OTR,Osmolarity,Nutrient Feeds,Cell Viability,Cell CountSelection StepsDevelopment StepsScaling a Process Requires Information.Scaling Steps Require Control.Selection Steps,Ap
7、proach:1.Nutrients,keep media the same and hope for the best.2.Oxygen Transfer Rate,keep mixing the same and hope for the best.3.pH,use buffers to control the process you get the idea.4.Temperature.Process Development Steps,Approach:1.Nutrients,keep constant-control media composition,feed timing and
8、 rates.2.Oxygen Transfer Rate,keep OTR the same,control agitator speeds and sparge gas compositions.3.pH,dual sided pH controller,use the same acid or base at each scale.4.Temperature,closed loop control over temperature.5.pCO2,Osmolarity,Cell Counts and Viabilities monitored,controls may be adjuste
9、d to match performance of previous scales.Selection Steps must be predictive of the first controlled scale.Development Steps constrain the culture to achieve reproducible results.Well plates,Shake Flasks,STBRsLarge Flasks/SpinnersTraditional Process Flow Upstream process development steps typically
10、rely on antiquated technologies.These technologies have questionable scalability.?Better Tools,Better Decisions,Earlier-FlasksSensored -FlasksSTBRs Improved consistency,monitoring and control permit better quality experiments.Improved experiment quality upstream of STBRs allows better decisions.-Fla
11、sks,SDR,SFR,AFS and The RAM make your existing work more predictive.AFM,The RAMu-FlaskThe RAMu-Flask+SDR,SFRAFS(Alcohol Fermentation System)ConsistencyMonitoringControl(limited)Consistency,Monitoring and Control-Flasks Test Tubes,Shake flasks.Increased aeration.Reduced evaporative loss.Solid support
12、,Re-useable/Autoclavable.May be used with the SensorDish Reader.-Flasks are Shake Flasks-Flasks Scale to Shake Flasks-Flask ConfigurationsFrom 100ul to 4.0mlApplikon Small-ScaleA big STEP AHEAD with a small footprint.-FlasksThe SDR and SFRPredictive Results,Earlier-Flask:Replaces well plate lids,tes
13、t tubes and shake flasks Works with all shaker-incubatorsSensorDish Reader+SensorFlask Reader Add Sensors to well plates and Shake Flasks Compatible with-flask closures Collect data from hundreds of parallel culturesSensorDish Reader,SDRSensorDish Reader,SDR1.Non-invasive,online.2.pH or Dissolved Ox
14、ygen.3.No Calibration Required.6-well24-wellExpandableAdditional SDRs link into the Basic Set.Up to 10 SDRs on one Laptop.No additional software required for expansion.SDRs Integrate with-Flasks-Flasks Integrate with the SensorDishReader1.Eliminate Well-plate inconsistency.2.Reduce and equalize evap
15、orative losses across well-plates.3.Obtain online pH or dissolved oxygen data for all wells.DO profiles(all 24 for each well-plate)Polystyreneu-FlaskSensorFlaskReader,SFRThe SensorFlask Reader,SFR Shaker-mounting blue-tooth sensor platform.Shake Flasks with DO and pH sensors.(also single DO or pH if
16、 desired)Multiple volumes(125/250/500/1000 ml).Up to 9 flasks on the platform at one time.Microbial and Mammalian ApplicationsMicrobial Applications:Monitor pH and DO levels.Ensure aerobic growth.Follow carbon source utilization and exhaustion.Adjust orbiter speeds to maintain DO levels for better s
17、calability.Mammalian Applications:Accurately monitor pH/pCO2 levels.Adjust bicarbonate levels more accurately.Monitor DO levels,adjust orbiter speeds for better scalability.SoftwareIndividual Flask CalibrationComplete Hardware Hardware Includes:Platform.Batteries.Bluetooth adapter.Software.Clamps.Bl
18、uetooth range of 15ft.Battery life 3 months on a single charge.(platform can operate on single battery)Predictive Results,Earlier-Flask:Replaces well plate lids,test tubes and shake flasks Works with all shaker-incubatorsSensorDish Reader+SensorFlask Reader Add Sensors to well plates and Shake Flasks Compatible with-flask closures Collect data from hundreds of parallel culturesQuestions?
