s-sepharose c-101 andy hunter alec mackenzie trevor seelert
TRANSCRIPT
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S-SepharoseC-101Andy HunterAlec MackenzieTrevor Seelert
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Purpose of UnitSeparate Proinsulin and other
proteins from basic impuritiesPrepare the stream for further
downstream purification
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Cation exchange Chromatography
EquilibrateLoadRinseEluteWashRegenerate
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Process Flow Sheet
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Process Flow Sheet
NewDesign
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Process Flow Sheet
NewDesign
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S-Sepharose DesignScale-up keeping constant values
Constant Parameter
Vairable Parameters
Column Height Column diameter
Residence time Volumetric flow rates
Linear velocities Amount of eluent
Rinse bed volume
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S-Sepharose Design
Parameter Lab Scale Process Scale
Column Height 0.8 m 0.8 m
Residence time 0.376991h 0.376991h
Load linear velocity 212.207 cm/h 212.207 cm/h
Eluent linear velocity 11 936.62 cm/h 11 936.62 cm/h
Column Diameter 0.1 m 1.574 m
Load Volumetric Flowrate
N/A 4.1288 m3/h
Scaled-up from US 101013 Patent
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S-Sepharose Design
GE Chromaflow GE AxiChrom
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S-Sepharose DesignGE Chromaflow ColumnCurrently max D = 1200mm, H =
30cmCustom Design available for D >
1200mm
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Economics and Suppliers
Unit Component Unit Cost
S-Sepharose $1000/L
Annual Report 601.35 L/yr
Total $601 350/yr
Chromatography column $241 000
Other plant direct costs $488 000
First Year TOTAL $1330350
S-Sepharose available from GE Healthcare
Chromaflow supplied by GE
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IEC AlternativesSP-Sepharose: Sulphopropyl
–O–CH2CHOHCH2OCH2CH2CH2SO3–
S-Sepharose: Sulphoethyl no longer produced –O–CH2–CH2–SO3–
Replaced by SP-SepharoseAnion exchange resin
Q-sepharose DEAE
HIC ColumnsSEC/Gel Filtration
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IEC Alternatives
DEAE: in accordance with German Patent 26 29 568
S-sepharose: in accordance with US Patent 5 101 013
Exchanger DEAE anion exchanger S-Sepharose cation exchanger
Matrix Vinyl polymer Crosslinked agarose gel
Buffer, gradient pH 9, 0.1-0.25 M NaCl pH 3.5, 0.15-0.35M NaCl
Loading 1 g/l 3 g/l
Yield 60% 80%
Purity 80-85% 90-95%
Separation Incomplete separation Base line separation
Comparison of IEC
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No Questions Allowed
Disclaimer: The views presented in this slide are not necessarily those of any or all team members.
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Just Kidding!