Purification of biomolecules combining ATPS and membrane chromatography

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

Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

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.

Originalspracheenglisch
Seiten (von - bis)152-160
Seitenumfang9
FachzeitschriftFood and Bioproducts Processing
Jahrgang92
Ausgabenummer2
DOIs
PublikationsstatusVeröffentlicht - 2014

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Biomolecules
Chromatography
Purification
chromatography
Membranes
ion exchange
bovine serum albumin
polyethylene glycol
Proteins
proteins
water
biopharmaceuticals
Ion exchange
Ion Exchange
hydrophobic bonding
Staphylococcal Protein A
binding capacity
Bovine Serum Albumin
salt concentration
Polyethylene glycols

Schlagwörter

    ASJC Scopus subject areas

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

    Dies zitieren

    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

    Purification of biomolecules combining ATPS and membrane chromatography. / Puthirasigamany, Mayuratheepan; Hamm, Ines; Van Winssen, Fatma Alexia; Nikbin, Nima; Kreis, Peter; Gorak, Andrzej; Zeiner, Tim.

    in: Food and Bioproducts Processing, Jahrgang 92, Nr. 2, 2014, S. 152-160.

    Publikation: Beitrag in einer FachzeitschriftArtikelForschungBegutachtung

    Puthirasigamany, M, Hamm, I, Van Winssen, FA, Nikbin, N, Kreis, P, Gorak, A & Zeiner, T 2014, 'Purification of biomolecules combining ATPS and membrane chromatography' Food and Bioproducts Processing, Jg. 92, Nr. 2, S. 152-160. https://doi.org/10.1016/j.fbp.2014.03.006
    Puthirasigamany M, Hamm I, Van Winssen FA, Nikbin N, Kreis P, Gorak A et al. Purification of biomolecules combining ATPS and membrane chromatography. Food and Bioproducts Processing. 2014;92(2):152-160. https://doi.org/10.1016/j.fbp.2014.03.006
    Puthirasigamany, Mayuratheepan ; Hamm, Ines ; Van Winssen, Fatma Alexia ; Nikbin, Nima ; Kreis, Peter ; Gorak, Andrzej ; Zeiner, Tim. / Purification of biomolecules combining ATPS and membrane chromatography. in: Food and Bioproducts Processing. 2014 ; Jahrgang 92, Nr. 2. S. 152-160.
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