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Purification of biomolecules combining ATPS and membrane chromatography

Paper ID Volume ID Publish Year Pages File Format Full-Text
18987 43040 2014 9 PDF Available
Title
Purification of biomolecules combining ATPS and membrane chromatography
Abstract

•Hybrid process: aqueous two phase extraction and membrane chromatography.•Influence of PEG and phosphate concentration on membrane chromatography.•Product recovery up to 99.9%.

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.

Keywords
Membrane chromatography; Aqueous two-phase extraction; Downstream processing
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Publisher
Database: Elsevier - ScienceDirect
Journal: Food and Bioproducts Processing - Volume 92, Issue 2, April 2014, Pages 152–160
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us