Purification of biomolecules combining ATPS and membrane chromatography

Mayuratheepan Puthirasigamany, Ines Hamm, Fatma Alexia Van Winssen, Nima Nikbin, Peter Kreis, Andrzej Gorak, Tim Zeiner*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Upstream processes for production of therapeutic proteins have been innovated and fermentation processes have been adopted for the use of recombinant microorganisms with high expression, but the downstream process is still the bottleneck in the biotechnological manufacturing process. A combined process consisting of aqueous two phase extraction (ATPE) and membrane chromatography is suggested to debottleneck downstream processing. ATPE has a large capacity, but the yield of the target product is from 74% to 97%. For this reason the product of ATPE waste stream is captured by membrane chromatography. In this work the binding capacity for the protein on protein A, ion exchange and hydrophobic exchange membrane chromatography was investigated experimentally with different concentration of polyethylene glycol (PEG), salt and protein. Protein A membrane was loaded with solutions resembling waste streams of ATPE for purifying IgG. For ion exchange and hydrophobic interaction membrane chromatography, the membrane was loaded with bovine serum albumin (BSA). PEG shows no significant effect on stability and capacity of membrane process. Even for small amount of BSA/IgG and high salt concentrations membrane adsorption is applicable. In this work it is demonstrated experimentally that a total product recovery of 99.9% for the purification of monoclonal antibody is possible.

Original languageEnglish
Pages (from-to)152-160
Number of pages9
JournalFood and Bioproducts Processing
Volume92
Issue number2
DOIs
Publication statusPublished - 2014

Keywords

  • Aqueous two-phase extraction
  • Downstream processing
  • Membrane chromatography

ASJC Scopus subject areas

  • Food Science
  • Biochemistry
  • Biotechnology
  • Chemical Engineering(all)

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  • Cite this

    Puthirasigamany, M., Hamm, I., Van Winssen, F. A., Nikbin, N., Kreis, P., Gorak, A., & Zeiner, T. (2014). Purification of biomolecules combining ATPS and membrane chromatography. Food and Bioproducts Processing, 92(2), 152-160. https://doi.org/10.1016/j.fbp.2014.03.006