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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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A new multicolumn, open-loop process for center-cut separation by solvent-gradient chromatography.

Ricardo J S Silva1, Rui C R Rodrigues, Hector Osuna-Sanchez

  • 1Requimte/CQFB, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.

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Summary

The new Gradient with Steady State Recycle (GSSR) process enables efficient center-cut separation for bioproducts. This chromatography method achieves high purity and yield, simplifying complex separations.

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Area of Science:

  • Chromatography
  • Bioseparations
  • Process Engineering

Background:

  • Center-cut separation is crucial for purifying bioproducts.
  • Existing methods can be complex and less efficient for ternary mixtures.
  • There is a need for versatile and easy-to-operate separation techniques.

Purpose of the Study:

  • To introduce and describe the Gradient with Steady State Recycle (GSSR) process.
  • To demonstrate the GSSR process's suitability for ternary separations, particularly for bioproducts.
  • To validate the GSSR process experimentally in a pilot unit.

Main Methods:

  • The GSSR process utilizes a multicolumn, open-loop system with cyclic steady-state operation.
  • It simulates a countercurrent solvent gradient with respect to the solid phase.
  • The process involves dynamic adjustment of the switching interval for optimal separation.

Main Results:

  • Experimental validation was performed using a crude peptide mixture purification via reversed-phase chromatography.
  • The GSSR process achieved high product purity (98%) and recovery (95%).
  • Stable operation was maintained in the pilot unit, demonstrating process robustness.

Conclusions:

  • The GSSR process is a versatile, flexible, and easy-to-operate method for center-cut separation.
  • It is particularly well-suited for the ternary separation of bioproducts, yielding three distinct fractions.
  • The experimental results confirm the GSSR process's effectiveness in achieving high purity and yield for target products.