Plasma fractionation using Simulated Moving Bed chromatography · Simulated Moving Bed...
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Plasma fractionation using Simulated Moving Bed chromatography F. Sander, M. Lübbert, Knauer Germany K. McCann, J. Bertolini, CSL Behring Australia PPB Meeting, Lanzarote 2013 16.05.2013
Transcript of Plasma fractionation using Simulated Moving Bed chromatography · Simulated Moving Bed...
Plasma fractionation using Simulated Moving Bed
chromatography
F. Sander, M. Lübbert, Knauer Germany K. McCann, J. Bertolini, CSL Behring Australia
PPB Meeting, Lanzarote 2013 16.05.2013
Outline
Why continuous chromatography? What is continuous chromatography? How does SMB work? What about Bio-applications? SMB in Plasma fractionation? What comes next?
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Why continuous chromatography?
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Why continuous chromatography?
What do we do normally?
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Why continuous chromatography?
Very high consumption of mobile and solid phase Long column or small particle sizes Low recovery rates and purities High dilution of the target compound
Column bed is not efficiently utilized Process is discrete and interrupted
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What is continuous chromatography?
SMB Binary or pseudo-binary mixtures Isocratic conditions
Most efficient chromatographic separation technique
MCSGP (Multi Column Countercurrent Solvent Gradient
Purification) Also for multi-component mixtures Using gradients for elution steps
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How does SMB work?
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Zone 3
Zone 1
Zon
e 2
Zon
e 4
Feed
Eluent Extract
Raffinate
How does SMB work?
Simulated Moving Bed Chromatography
Simulated countercurrent movement of solid and liquid phase Continuous column switching No dilution
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Column switching
Cyclic Steady State
Eluent Extract Feed Raffinate
How does SMB work? Outlet Profiles
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What are the advantages?
Higher purities and yields Decreased solvent consumption Cheaper column materials No dilution No interruption
Highly efficient process! 16/05/2013 Sander, PPBM 2013 10
What are typical applications?
Petrochemistry Sorbex Process (UOP)
Food processing Citric acid Sugar separations
Pharmaceuticals Chiral separations
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What about Bio-applications?
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What about Bio-applications?
Limitations Multi-component mixtures Two step chromatography Column regeneration
But in batch
High product loss High dilution of product High buffer consumption
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Transfer to SMB
Device CSEP C9116 Biocompatible SMB
Columns Special glass columns Self-fillable under pressure
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SMB in Plasma fractionation?
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SMB in Plasma fractionation?
Current method for chromatographic purification of albumin
Precipitation Delipidation Chromatographic methods
– Anion exchange
– Cation exchange
– Size exclusion
Aim Purification of HSA from plasma
SMB???
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What was the first step? Stationary phase
Weak anion exchanger (BioFox40 DEAE)
Column
Bioline Glass column Liquid phase
Acetate buffer, pH 5.5 Flow rate
2 mL/min
Globulin fraction
HSA
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What was the second step?
8 columns Isocratic separation
Process simulation:
Feed flow: 0.17 mL/min Eluent flow: 1.64 mL/min
Unpurified Feed
Purified Extract
HSA fraction
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Results Unpurified Feed
Purified Extract
HSA fraction
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Feed purity: 85% Extract purity: 98%
No Albumin in purified raffinate
Very promising method!
Results
Low levels of transferrin and IgG in the raffinate
Accumulation on the column? Purging with 1 M NaCl and NaOH
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What comes next? Further process optimization
Improvement of the AIEX step Polishing step
Transferring SEC polishing step to SMB
MCSGP
Comparing results
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Acknowledgement CSL Behring, Australia
Karl McCann Joseph Bertolini
My colleagues at Knauer
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