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Design of RP-HPLC 1 for Insulin Production

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Design of RP-HPLC 1 for Insulin Production Powered By Docstoc
					   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
     Particlesize (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 Å
                  Diameter 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!

				
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posted:4/8/2010
language:English
pages:17