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Design of RP-HPLC 1 for Insulin Production

Design of RP-HPLC 1 for Insulin Production. Presented by: Patricia Yee, Nicholas Irianto, Thomas Fang. Overview. Background Information Significance of RP-HPLC1 Design Parameters Selected Design and Specifications Questions. Background Information. What is HPLC ?

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Design of RP-HPLC 1 for Insulin Production

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  1. Design of RP-HPLC 1 for Insulin Production Presented by: Patricia Yee, Nicholas Irianto, Thomas Fang

  2. Overview • Background Information • Significance of RP-HPLC1 • Design Parameters • Selected Design and Specifications • Questions

  3. Background Information • What is HPLC? • High Performance Liquid Chromatography - or - • High-Pressure Liquid Chromatography

  4. How HPLC works…

  5. Reverse-Phase HPLC

  6. Insulin Production Process:RP-HPLC 1

  7. Insulin Production Process:RP-HPLC 1 • What comes out… 1) Contaminated protein, denatured insulin, enzyme and small amounts of insulin ester and WFI 2) Majority of desired insulin ester and WFI

  8. Insulin Production Process:RP-HPLC 1 • Gradient Elution of mobile phase: • Acetonitrile / Water / Acid

  9. Main Design Parameters • Column • Size (Internal diameter) • Material (Stainless Steels, Glass, etc)

  10. Main Design Parameters • Resin • Particle size (10μm ~75μm ) • Pore size (300 Á) • Material (Silica, styrene-divinylbenzene, etc.)

  11. Other Design Considerations • Pump (pressure capacity) • Solvent • Acetonitrile • Polar solvent (increase retention time) • Hydrophobic solvent (decrease retention time)

  12. Possible Design

  13. Design Specifications • 100% binding and 90% yield for insulin ester • 60% binding and 5% yield for denatured insulin precursors • Purification results in outlet stream consisting of 340 kg insulin ester/batch

  14. Design Selections • HPLC Columns • Novasep Prochrom® DAC Columns LC1600 • H x D: 450 x 1600 mm • HPLC Resins • PRP-3 standard Poly(styrene-divinylbenzene) (PSDVB) • Pore size 300 Å • Diameter 10 μm

  15. Design Justifications • Stable from pH 1-13 • Does not contain acidic silanol groups, which are present on silica supports. • Highly cross-linked for exceptional durability and to prevent shrinking and • Consistent performance • 10μm particle diameter is selected for better solution and peak shape

  16. Design Details • Twenty three LC 1600 HPLC column units with 0.91 hours process time would be required for the process according to SuperPro • For one LC 1600 HPLC column with maximum bed volume of 900 L would require 360 000 grams of PRP-3 resins COSTS • $24/g of PRP-3 resins This translates to $ 8 640 000 / LC 1600 HPLC column For 23 HPLC columns, it would cost $ 198 720 000 • $28 000 for a Novasep Prochrom DAC LC 1600 column Total Cost $ 200 000 000

  17. Questions? THANK YOU!

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