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S-Sepharose C-101 Andy Hunter Alec Mackenzie Trevor Seelert.

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Presentation on theme: "S-Sepharose C-101 Andy Hunter Alec Mackenzie Trevor Seelert."— Presentation transcript:

1 S-Sepharose C-101 Andy Hunter Alec Mackenzie Trevor Seelert

2 Purpose of Unit Separate Proinsulin and other proteins from basic impurities Prepare the stream for further downstream purification

3 Cation exchange Chromatography Equilibrate Load Rinse Elute Wash Regenerate

4 Process Flow Sheet

5 New Design

6 Process Flow Sheet New Design

7 S-Sepharose Design Scale-up keeping constant values Constant ParameterVairable Parameters Column Height Column diameter Residence time Volumetric flow rates Linear velocities Amount of eluent Rinse bed volume

8 S-Sepharose Design ParameterLab ScaleProcess Scale Column Height 0.8 m Residence time 0.376991h Load linear velocity 212.207 cm/h Eluent linear velocity 11 936.62 cm/h Column Diameter 0.1 m1.574 m Load Volumetric Flowrate N/A4.1288 m3/h Scaled-up from US 101013 Patent

9 S-Sepharose Design GE Chromaflow GE AxiChrom

10 S-Sepharose Design GE Chromaflow Column Currently max D = 1200mm, H = 30cm Custom Design available for D > 1200mm

11 Economics and Suppliers Unit ComponentUnit Cost S-Sepharose$1000/L Annual Report601.35 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

12 IEC Alternatives SP-Sepharose: Sulphopropyl  –O–CH 2 CHOHCH 2 OCH 2 CH 2 CH 2 SO 3 –  S-Sepharose: Sulphoethyl  no longer produced  –O–CH 2 –CH 2 –SO 3 –  Replaced by SP-Sepharose Anion exchange resin  Q-sepharose  DEAE HIC Columns SEC/Gel Filtration

13 IEC Alternatives DEAE: in accordance with German Patent 26 29 568 S-sepharose: in accordance with US Patent 5 101 013 ExchangerDEAE anion exchanger S-Sepharose cation exchanger MatrixVinyl polymerCrosslinked agarose gel Buffer, gradientpH 9, 0.1-0.25 M NaClpH 3.5, 0.15-0.35M NaCl Loading1 g/l3 g/l Yield60%80% Purity80-85%90-95% SeparationIncomplete separationBase line separation Comparison of IEC

14 No Questions Allowed Disclaimer: The views presented in this slide are not necessarily those of any or all team members.

15 Just Kidding!

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