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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

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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


April 02 2008 yannick beauregard queenie chow irena doslo

Intelligen., 1991


Diafiltration unit df 101

Diafiltration unit (df-101)

Intelligen., 1991


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


Diafiltration volume superpro modifications

Diafiltration volume (SuperPro modifications)


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)


April 02 2008 yannick beauregard queenie chow irena doslo

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)


April 02 2008 yannick beauregard queenie chow irena doslo

QUESTIONS?

Thank you!


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