BACKGROUND: The use of antibodies, such as immunoglobulin G (IgG), has faced a significant growth in the past decades for biomedical and research purposes. However, antibodies are high cost biopharmaceuticals, for which the development of alternative and cost-effective purification strategies is still in high demand. RESULTS: Aqueous biphasic systems (ABS) composed of poly(propylene glycol) (PPG) and cholinium-based ionic liquids (ILs) were investigated for the separation of IgG. The ABS phase diagrams were determined and characterized whenever required. Initial optimization studies with commercial IgG were carried out, followed by the extraction of IgG from rabbit serum. In all ABS, IgG preferentially partitions to the IL-rich phase, unveiling preferential interactions between IgG and ILs. Good results were obtained with commercial IgG, with extraction efficiencies ranging between 93% and 100%, and recovery yields ranging between 20% and 100%. Two of the best and two of the worst identified ABS were then evaluated in what concerns their performance to separate and recover IgG from rabbit serum. With these ABS, extraction efficiencies of 100% and recovery yields > 80% were obtained, indicating an increase in the recovery yield and extraction efficiencies when using real matrices. Under the best conditions studied, IgG with a purity level of 49% was obtained in a single-step. This purity level of IgG is higher than those previously reported using other IL-polymer ABS. CONCLUSION: IgG preferentially migrates to the IL-rich phase in ABS formed by ILs and polymers, allowing the design of effective separation systems for its recovery from serum samples.
BACKGROUND: The use of antibodies, such as immunoglobulin G (IgG), has faced a significant growth in the past decades for biomedical and research purposes. However, antibodies are high cost biopharmaceuticals, for which the development of alternative and cost-effective purification strategies is still in high demand. RESULTS: Aqueous biphasic systems (ABS) composed of poly(propylene glycol) (PPG) and cholinium-based ionic liquids (ILs) were investigated for the separation of IgG. The ABS phase diagrams were determined and characterized whenever required. Initial optimization studies with commercial IgG were carried out, followed by the extraction of IgG from rabbit serum. In all ABS, IgG preferentially partitions to the IL-rich phase, unveiling preferential interactions between IgG and ILs. Good results were obtained with commercial IgG, with extraction efficiencies ranging between 93% and 100%, and recovery yields ranging between 20% and 100%. Two of the best and two of the worst identified ABS were then evaluated in what concerns their performance to separate and recover IgG from rabbit serum. With these ABS, extraction efficiencies of 100% and recovery yields > 80% were obtained, indicating an increase in the recovery yield and extraction efficiencies when using real matrices. Under the best conditions studied, IgG with a purity level of 49% was obtained in a single-step. This purity level of IgG is higher than those previously reported using other IL-polymer ABS. CONCLUSION: IgG preferentially migrates to the IL-rich phase in ABS formed by ILs and polymers, allowing the design of effective separation systems for its recovery from serum samples.
Authors: Martyn J Earle; José M S S Esperança; Manuela A Gilea; José N Canongia Lopes; Luís P N Rebelo; Joseph W Magee; Kenneth R Seddon; Jason A Widegren Journal: Nature Date: 2006-02-16 Impact factor: 49.962
Authors: Maria V Quental; Magda Caban; Matheus M Pereira; Piotr Stepnowski; João A P Coutinho; Mara G Freire Journal: Biotechnol J Date: 2015-04-30 Impact factor: 4.677
Authors: Mara G Freire; Jorge F B Pereira; María Francisco; Héctor Rodríguez; Luís Paulo N Rebelo; Robin D Rogers; João A P Coutinho Journal: Chemistry Date: 2012-01-03 Impact factor: 5.236
Authors: Cláudia L S Louros; Ana Filipa M Cláudio; Catarina M S S Neves; Mara G Freire; Isabel M Marrucho; Jérôme Pauly; João A P Coutinho Journal: Int J Mol Sci Date: 2010-04-15 Impact factor: 5.923