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April 02 2008 Yannick Beauregard Queenie Chow Irena Doslo. Diafiltration unit (DF-101) in Monoclonal antibody production . Background on diafiltration.

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april 02 2008 yannick beauregard queenie chow irena doslo

April 02 2008Yannick BeauregardQueenie ChowIrena Doslo

Diafiltration unit (DF-101) in Monoclonal antibody production

background on diafiltration
Background on diafiltration
  • Is a technique that uses basic principles of filtration to completely remove, replace or lower the concentration of salts or solvents from solutions containing biomolecules
  • Uses permeable membrane to separate the components mainly based on size
  • Dilution and concentration
  • Dialysis, column-based gel filtration
diafiltration
Diafiltration

Millipore Inc., 2003

design considerations
Design Considerations
  • Type of flow (tangential vs. direct)
  • Membrane selection
  • Type of diafilter modules
  • Diafiltration volumes
  • Continuous vs. discontinuous flow
tangential vs direct flow
Tangential vs. direct flow
  • Direct Flow:
  • Large molecule trapped on membrane and forms gel
  • More susceptible to fouling
  • Flux rate decreases as volume filtered increases

Millipore Inc., 2003

  • Tangential Flow
  • Solute diffuses through the surface of the membrane tangent to the flow of the feed
  • Minimize build up of molecules – less fouling
  • Prevents rapid decline in flux rate

Millipore Inc., 2003

membrane selection criteria
Membrane selection criteria
  • Primarily based on size of biomolecule
  • Molecular weight cut off (MWCO) of the membrane should be 1/3rdto 1/5th of the MW of the molecule to be retained
    • Typical MW of mAb: 150kDa => 30000 MWCO
  • Other considerations- surface chemistry
  • Membrane flux rate- time factor vs. product recovery
    • For protein separation: 30 LMH
types of diafilters
Types of diafilters
  • Flat sheet tangential flow
  • Hollow fibre
  • Tubular
  • Spiral wound

Millipore Inc., 2003

continuous vs discontinuous filtration
Continuous vs. discontinuous filtration
  • Continuous
    • Typically constant volume
      • Removal rate of salt = addition rate of water
      • Not the case in SuperPro model
      • Addition of WFI is at 1/3rd of the removal rate of salt (filtrate) in SuperPro
    • More suited for process scale- requires pumps
  • Discontinuous
    • Concentration and dilution cycles
    • Usually more feasible on a laboratory scale
final design
Final design
  • Hollow fiber cartridges
  • Membrane area needed
    • Mem. area = filtrate vol / (filtrate flux * process time)
    • 22.38 m2 with a 5% design consideration
  • From GE Healthcare services
    • ProCell ultrafiltration unit with 30000 MWCO pore size and seven 3.7 m2 membranes
      • UNIT NUMBER:
    • Stainless stain housing
    • Steam in place cartridges can be added

GE Healthcare (2007)

GE Healthcare (2007)

slide13
Cost
  • Capital cost
    • Equipments
      • Membrane holder
      • Pumps, valves, piping
      • Type of instrumentation
  • Material cost
    • Membrane area
    • Water and chemical usage
  • Labour cost
    • Manual or automatic
    • In SuperPro, the design unit with 3 DV is $56000

GE Healthcare (2007)

slide14

QUESTIONS?

Thank you!

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