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? High Performance Liquid Chromatography

- or - High-Pressure Liquid Chromatography

How HPLC works…

Reverse-Phase HPLC

Insulin Production Process: RP-HPLC 1

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

Insulin Production Process: RP-HPLC 1

Gradient Elution of mobile phase: Acetonitrile / Water / Acid

Main Design Parameters Column

Size (Internal diameter)Material (Stainless Steels, Glass, etc)

Main Design Parameters Resin

Particle size (10μm ~75μm )Pore size (300 Á)Material (Silica, styrene-divinylbenzene,

etc.)

Other Design Considerations

Pump (pressure capacity) Solvent

Acetonitrile Polar solvent (increase retention time) Hydrophobic solvent (decrease retention

time)

Possible Design

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

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 ÅDiameter 10 μm

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

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

Questions?

THANK YOU!