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Anion Exchange Column

Anion Exchange Column. By: Eilish McConville Justin Low. Presentation Overview. Theory and Background Column Sizing Timing Engineering Design Costs Recommendations. Theory and Background. Chromatography separation method differentiating species based on the ionic charge Four Stages:

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Anion Exchange Column

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  1. Anion Exchange Column By: Eilish McConville Justin Low

  2. Presentation Overview • Theory and Background • Column Sizing • Timing • Engineering Design • Costs • Recommendations

  3. Theory and Background • Chromatography separation method differentiating species based on the ionic charge • Four Stages: • Initial Binding • Wash • Elute • Rebuffer • Elution Profile

  4. Column Sizing • Calculated Protein Loading vs. Volume • Used Empirical data from Bio-Rad • Model: Protein Load (mg) = 15.007*Volume(mL)-0.125

  5. Column Sizing • Pick Suitable Column • Consider Size, efficiency and cost • GE Chromaflow column selected • Volume = 942.5 L • Calculate Total Number of Column Runs for single column • Total amount of protein/batch = 202.80kg • Can separate 7.071kg/column run • Need to run column 28.67 times

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

  7. Number of Columns • Four columns is ideal • Operation can be offset to allow for continuous product

  8. Engineering Design

  9. Design Considerations • Insulin Ester Isoelectric Point = 5.3 • Bead Choice: • DEAE-Sephadex A-25 • Medium to coarse beads (170-260 µm) • Initial buffer pH • Flow rate chosen for four columns • 50% Protein Saturation

  10. Cost • GE quoted price = $150,577 (CAN)/column • Total Capital Cost ~ $900 000 (CAN) • Amortized Cost (20 years, 10%) ~ $100 000/year • Operating Costs • Gel Matrix refills (~$2000/kg or $2.6M/column) • Utility costs • Wash and Eluent Material Costs • Maintenance • Other Capital Costs not considered • Pumps • Holding Tanks • Control System

  11. Manufacturers • GE • Bio-Rad Laboratories • Dionex Corp. • Shodex Inc. • Thermo Labs • Sartorius AG • Sigma Aldrich • Varian Inc. • UpFront Chromatography

  12. Recommendations • Experimentation with the unit to determine: • Optimal buffer pH • Protein loading • Talk with suppliers: • General Electric Company • UpFront Chromatography

  13. Questions

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