Design of RPHPLC 1 for Insulin Production

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!