TY - JOUR
T1 - Purification of biomolecules combining ATPS and membrane chromatography
AU - Puthirasigamany, Mayuratheepan
AU - Hamm, Ines
AU - Van Winssen, Fatma Alexia
AU - Nikbin, Nima
AU - Kreis, Peter
AU - Gorak, Andrzej
AU - Zeiner, Tim
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - Aqueous two-phase extraction
KW - Downstream processing
KW - Membrane chromatography
UR - http://www.scopus.com/inward/record.url?scp=84898971638&partnerID=8YFLogxK
U2 - 10.1016/j.fbp.2014.03.006
DO - 10.1016/j.fbp.2014.03.006
M3 - Article
AN - SCOPUS:84898971638
SN - 0960-3085
VL - 92
SP - 152
EP - 160
JO - Food and Bioproducts Processing
JF - Food and Bioproducts Processing
IS - 2
ER -