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Suspended bed chromatography, a new approach in downstream processing.

P R Levison1, A K Hopkins, P Hathi

  • 1Whatman International Ltd., Springfield Mill, Maidstone, Kent, UK. peter1@whatman.co.uk

Journal of Chromatography. A
|September 8, 2000
PubMed
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A novel suspended bed chromatography technique enhances downstream processing. This method combines batch adsorption benefits with enclosed column efficiency for faster protein purification.

Area of Science:

  • Biochemistry
  • Chemical Engineering
  • Separation Science

Background:

  • Downstream processing in biotechnology often involves complex and time-consuming purification steps.
  • Traditional batch adsorption methods can be inefficient and difficult to scale.
  • Existing column chromatography systems may have limitations in handling large volumes or specific adsorbent types.

Purpose of the Study:

  • To introduce and evaluate a new hybrid technique called suspended bed chromatography for protein purification.
  • To demonstrate the feasibility of using established contactors and adsorbents in this novel system.
  • To assess the efficiency and speed of suspended bed chromatography for purifying ovalbumin from hen-egg white.

Main Methods:

  • Utilized a 44 cm I.D. IsoPak column and Express-Ion Exchanger Q anion-exchange cellulose.

Related Experiment Videos

  • Employed the IsoPak slurry preparation station for adsorbent suspension and feedstock recirculation.
  • Performed protein adsorption, washing, and gradient elution desorption within the IsoPak column unit.

    • Main Results:

    • Successfully purified ovalbumin from hen-egg white using suspended bed chromatography.
    • The entire purification process, including adsorption, washing, and desorption, was completed in under 3 hours.
    • Achieved efficient protein loading and desorption using a flow rate of 300 cm/h.

    Conclusions:

    • Suspended bed chromatography offers a new, efficient approach to downstream processing.
    • This hybrid technique provides a less challenging alternative to traditional batch separations.
    • The method is compatible with existing equipment and adsorbents, facilitating its adoption.