title: Magnetic aqueous two phase fishing: a hybrid process technology for antibody purification.
abstract: The potential to combine aqueous two-phase extraction (ATPE) with magnetic separation was here investigated with the aim of developing a selective non-chromatographic method for the purification of antibodies from cell culture supernatants. Aqueous two-phase systems (ATPS) composed of polyethylene glycol (PEG) and dextran were supplemented with several surface modified magnetic particles (MPs) at distinct salt concentrations. The partition of pure human IgG in the upper and lower phases as well as the amount adsorbed at the MPs surface was investigated, indicating that MPs coated with dextran and gum Arabic established the lowest amount of non-specific interactions. The binding capacity of gum arabic coated particles modified with aminophenyl boronic acid (GA-APBA-MP) was were found to be excellent in combination with the ATPS system, yielding high yields of antibody recovery (92%) and purity (98%) from cell culture supernatants. The presence of MPs in the ATPS was found to speed up phase separation (from 40 to 25min), to consume a lower amount of MPs (half of the amount needed in magnetic fishing) and to increase the yield and purity of a mAb purified from a cell culture supernatant, when compared with ATPE or magnetic fishing processes alone.
authors.affiliation: REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Vijaykumar L
authors.initials: VL
authors.lastname: Dhadge
authors.affiliation: Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisbon, Portugal.
authors.forename: Sara A S L
authors.initials: SA
authors.lastname: Rosa
authors.affiliation: Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisbon, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Azevedo
authors.affiliation: Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisbon, Portugal.
authors.forename: Raquel
authors.initials: R
authors.lastname: Aires-Barros
authors.affiliation: REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal. Electronic address: cecilia.roque@fct.unl.pt.
authors.forename: Ana C A
authors.initials: AC
authors.lastname: Roque
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001897:Boronic Acids; D016466:CHO Cells; D003412:Cricetulus; D003911:Dextrans / Q000737:chemistry*; D005240:Feasibility Studies; D005290:Ferric Compounds / Q000737:chemistry*; D006170:Gum Arabic; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D060328:Magnetic Phenomena; D059346:Magnets; D011092:Polyethylene Glycols / Q000737:chemistry*; D014867:Water
chemical_list: D000911:Antibodies, Monoclonal; D001897:Boronic Acids; D003911:Dextrans; D005290:Ferric Compounds; D007074:Immunoglobulin G; D014867:Water; C000499:ferric oxide; C028592:3-aminobenzeneboronic acid; D011092:Polyethylene Glycols; D006170:Gum Arabic
keywords: Antibodies; Aqueous two-phase system; Boronic acid; Hybrid process; Magnetic particle