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

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Presentation on theme: "Design of RP-HPLC 1 for Insulin Production Presented by: Patricia Yee, Nicholas Irianto, Thomas Fang."— Presentation transcript:

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 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 grams of PRP-3 resins COSTS $24/g of PRP-3 resins This translates to $ / LC 1600 HPLC column For 23 HPLC columns, it would cost $ $ for a Novasep Prochrom DAC LC 1600 column Total Cost $

17 Questions? THANK YOU!


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