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Sample Displacement Batch Chromatography of Proteins.

Laura Heikaus1, Siti Hidayah1, Manasia Gaikwad1

  • 1Institute of Clinical Chemistry and Laboratory Medicine, Mass Spectrometric Proteomics Group, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

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|October 31, 2020
PubMed
Summary

This study presents fast, inexpensive methods for large-scale protein purification using sample displacement batch chromatography. These protocols simplify establishing preparative chromatography for biopharmaceutical manufacturing.

Keywords:
Anion exchange chromatographyCohn fractionPlasma proteinsPreparative chromatographySDS-PAGESample displacement chromatographySimulated-moving-bed

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Area of Science:

  • Biochemistry
  • Chemical Engineering
  • Biotechnology

Background:

  • Large-scale chromatography is crucial for downstream processing in biopharmaceutical development.
  • Screening experiments and pilot-plant chromatography are essential but can be time-consuming and costly.

Purpose of the Study:

  • To develop fast, simple, and inexpensive methods for establishing preparative chromatography.
  • To demonstrate these methods using anion-exchange chromatography of human plasma proteins.
  • To provide adaptable protocols for large-scale protein purification and biopharmaceutical manufacturing.

Main Methods:

  • Utilized sample displacement batch chromatography for protein mixture separation.
  • Performed anion-exchange chromatography on a human plasma protein fraction (Cohn IV-4).
  • Included screening experiments and upscaled chromatography by a factor of 100, monitored by SDS-PAGE.

Main Results:

  • Successfully established preparative chromatography methods for complex protein mixtures.
  • Demonstrated efficient screening and upscaling processes for large-scale purification.
  • Validated the methods through the separation of human plasma proteins.

Conclusions:

  • The developed protocols are easily adaptable for large-scale chromatographic purification of various proteins.
  • These methods are suitable for both laboratory settings and biopharmaceutical manufacturing.
  • Findings can inform the development of packed columns for simulated-moving-bed (SMB) chromatography.