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Anion Exchange Column By: Eilish McConville Justin Low.

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Presentation on theme: "Anion Exchange Column By: Eilish McConville Justin Low."— Presentation transcript:

1 Anion Exchange Column By: Eilish McConville Justin Low

2 Presentation Overview Theory and BackgroundTheory and Background Column SizingColumn Sizing TimingTiming Engineering DesignEngineering Design CostsCosts RecommendationsRecommendations

3 Theory and Background Chromatography separation method differentiating species based on the ionic chargeChromatography separation method differentiating species based on the ionic charge Four Stages:Four Stages: Initial BindingInitial Binding WashWash EluteElute RebufferRebuffer Elution ProfileElution Profile

4 Column Sizing Calculated Protein Loading vs. VolumeCalculated Protein Loading vs. Volume Used Empirical data from Bio-RadUsed Empirical data from Bio-Rad Model:Model: Protein Load (mg) = *Volume(mL)-0.125

5 Column Sizing Pick Suitable ColumnPick Suitable Column Consider Size, efficiency and costConsider Size, efficiency and cost GE Chromaflow column selectedGE Chromaflow column selected Volume = LVolume = L Calculate Total Number of Column Runs for single columnCalculate Total Number of Column Runs for single column Total amount of protein/batch = kgTotal amount of protein/batch = kg Can separate 7.071kg/column runCan separate 7.071kg/column run Need to run column timesNeed to run column times

6 Timing Need to consider two time frames:Need to consider two time frames: Batch Process time for columnBatch Process time for column Continuous process time for entire insulin productionContinuous process time for entire insulin production Process requires multiple columnsProcess requires multiple columns Chose a flow rate that allowed for four columnsChose a flow rate that allowed for four columns L/min (72% of maximum)93.82 L/min (72% of maximum) Determined the step times based on flow rateDetermined the step times based on flow rate

7 Number of Columns Four columns is idealFour columns is ideal Operation can be offset to allow for continuous productOperation can be offset to allow for continuous product Step Volume (L) Time (min) Insulin Ester solution Wash Elution Rebuffer Total

8 Engineering Design

9 Design Considerations Insulin Ester Isoelectric Point = 5.3Insulin Ester Isoelectric Point = 5.3 Bead Choice:Bead Choice: DEAE-Sephadex A-25DEAE-Sephadex A-25 Medium to coarse beads ( µ m)Medium to coarse beads ( µ m) Initial buffer pHInitial buffer pH Flow rate chosen for four columnsFlow rate chosen for four columns 50% Protein Saturation50% Protein Saturation

10 Cost GE quoted price = $150,577 (CAN)/columnGE quoted price = $150,577 (CAN)/column Total Capital Cost ~ $ (CAN)Total Capital Cost ~ $ (CAN) Amortized Cost (20 years, 10%) ~ $ /yearAmortized Cost (20 years, 10%) ~ $ /year Operating CostsOperating Costs Gel Matrix refills (~$2000/kg or $2.6M/column)Gel Matrix refills (~$2000/kg or $2.6M/column) Utility costsUtility costs Wash and Eluent Material CostsWash and Eluent Material Costs MaintenanceMaintenance Other Capital Costs not consideredOther Capital Costs not considered PumpsPumps Holding TanksHolding Tanks Control SystemControl System

11 Manufacturers GEGE Bio-Rad LaboratoriesBio-Rad Laboratories Dionex Corp.Dionex Corp. Shodex Inc.Shodex Inc. Thermo LabsThermo Labs Sartorius AGSartorius AG Sigma AldrichSigma Aldrich Varian Inc.Varian Inc. UpFront ChromatographyUpFront Chromatography

12 Recommendations Experimentation with the unit to determine:Experimentation with the unit to determine: Optimal buffer pHOptimal buffer pH Protein loadingProtein loading Talk with suppliers:Talk with suppliers: General Electric CompanyGeneral Electric Company UpFront ChromatographyUpFront Chromatography

13 Questions


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