Purification of human immunoglobulin G by thermoseparating aqueous two-phase systems

The purification of human immunoglobulin G (IgG) from a Chinese hamster ovary (CHO) cells supernatant was studied using an aqueous two-phase system (ATPS) composed of ethylene oxide/propylene oxide (UCON) and dextran. In UCON/dextran systems IgG partitions preferentially to the less hydrophobic dext...

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Bibliographic Details
Published in:Journal of Chromatography A 2008-06, Vol.1195 (1), p.94-100
Main Authors: Ferreira, I. Filipa, Azevedo, Ana M., Rosa, Paula A.J., Aires-Barros, M. Raquel
Format: Article
Language:English
Subjects:
Animals
Aqueous two-phase system
Biological and medical sciences
Biotechnology
Chemical Fractionation - methods
CHO Cells
Cricetinae
Cricetulus
Dextrans - chemistry
Epoxy Compounds - chemistry
Ethylene oxide
Ethylene Oxide - chemistry
Fundamental and applied biological sciences. Psychology
Humans
IgG
Immunoglobulin G - chemistry
Immunoglobulin G - isolation & purification
Ligand enhanced partitioning
Methods. Procedures. Technologies
Others
Propylene oxide
Temperature
Temperature sensitive polymers
Various methods and equipments
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Summary:The purification of human immunoglobulin G (IgG) from a Chinese hamster ovary (CHO) cells supernatant was studied using an aqueous two-phase system (ATPS) composed of ethylene oxide/propylene oxide (UCON) and dextran. In UCON/dextran systems IgG partitions preferentially to the less hydrophobic dextran-rich phase (Kp < 1). The addition of triethylene glycol-diglutaric acid (TEG-COOH) shifted the IgG partition into the upper phase showing significant improvements in both the recovery yields and purity. The purification of IgG from a CHO cell supernatant with UCON 2000/dextran/TEG-COOH system was optimised using a central composite design. Using an ATPS composed of 8% UCON, 6% dextran and 20% TEG-COOH, IgG was purified, in two steps, with a global yield of 85% and 88% purity. Statistical valid models were obtained to predict the effect of the experimental conditions on the IgG yield and purity, for both extraction and back-extraction steps. A system composed of 10% UCON, 5.5% dextran and 20% TEG-COOH was identified as the best compromise between final purity and yield.
ISSN:0021-9673
DOI:10.1016/j.chroma.2008.04.077