生物反应器课件 bioreactor design Select cell culture method

1、Bioreactor designSelect cell culture methodContent Introduction Cell culture specification Agitation Temperature control Sterilization CIPBioreactor design considerationsGoals: Max growth or max productivity Need stringently maintained process conditions Agitation (max transfer) Homogenous condition

2、Bioreactor design considerationsRepeatable transfer of energy, substrates, metabolites.- Good mixing depends on: nature medium rheology of the broth airflow rate/ type of aeration kind and number of impellers/baffles organism behaviour (shearstress) geometry of vessel (H/D) accurate speed controlBio

3、reactor design considerationsBioreactor design considerations for cell culture Top driven agitator Marine impellers Tipspeed up to 1 m/s No baffles Micro porous sparger Gas flows up to 0.1 vvm Headspace aeration Aspect ratio = 1 at working volume CIP-able designBioreactor design considerations Shape

4、 ASME Bottoms Hemispherical Bottoms Jacket Design Conventional Dimpled Half Pipe Ports Tri-Clamp, DN, NA-connect, Ingold Type, Flange typeBioreactor design considerationsASME Head(s)Hemispherical Head(s)Agitation Seals Mechanical Face Seals Lip Seals Magnetic Drives Impellers Turbine Marine Pitched

5、Blade OthersAgitationAgitation - turbine impeller For unaerated water 20 C in a fully baffled tank the following formula applies: P= 4.5 x 10-13 * Di5 * N3 * FDi = Impeller Diameter in InchN = RPM of ShaftF = One (1) for one impeller (1.8) for two impellers (2.4) for three impellersAgitation - turbi

6、ne impellerTip Speed should also be considered because of tendency to shear cells. Do not exceed 1 m/s for cell cultureAnd 5 - 6 m/s for microbial cultures.Temperature controlTemperature controlCooling System: remove 50 to 100 Watts/Liter of Volume.Heating Systems: One (1) degree C per minute betwee

7、n 25 and 45 CSpargersRing TypeSingle OrificeSinteredFiltersMembrane (0.2/0.45 micron pore size)Usually made of TeflonAdvantages - ValidatableDisadvantages - Relatively Expensive Blinded by WaterDepth CartridgesAdvantages - Not blinded by WaterRelatively efficientDisadvantages - Not easily validatedI

8、ntegrity testing: W.I.T. or bubble pointSterility Considerations Killing Rates Dead Legs Fitting Types Piping ConsiderationsSterility ConsiderationsCritical Elements of Steam Sterilization Time Temperature MoistureAir RemovalSterility ConsiderationsSterility Considerations Killing rate F0 = ti * 10(

9、Ti - 121) / Z F0 = Integrated amount of lethality delivered during a cyclei.e. A cycle with F0 = 17 minutes has a lethality equivalent to 17 minutes at121 C.CIP considerations Sterile Piping should be pitched for drainability. CIP FluidsDetergentAlkaline AcidWFI rinse Clean Steam Circuit for Distribution Sprayballs should be designed to deliver 30 LPM per meter of vessel circumference. Minimum flow rate of 1.5 meter per second for pipe or tubing for effective cleaning. EP surface finishingCIP Considera