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Purification of human antibodies from animal cell cultures using gum arabic coated magnetic particles
Author(s) -
Alves Bruno Martins,
Borlido Luís,
Rosa Sara A.S.L.,
Silva Marta F.F.,
AiresBarros Maria Raquel,
Roque Ana C.A.,
Azevedo Ana M.
Publication year - 2015
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.4378
Subject(s) - monoclonal antibody , elution , chromatography , chemistry , chinese hamster ovary cell , adsorption , antibody , yield (engineering) , expanded bed adsorption , biochemistry , biology , materials science , receptor , organic chemistry , immunology , metallurgy
BACKGROUND The emergence of monoclonal antibodies ( mAbs ) as new biopharmaceutical products requires the development of new purification methods that are not only effective but are able to reduce production costs. To address the problematic recovery of mAbs , gum arabic ( GA ) coated magnetic particles ( MPs ) were used for the purification of human antibodies from animal cells supernatants. RESULTS MPs were synthesized via co‐precipitation and exhibited a spherical‐like physical aspect, with an average hydrodynamic diameter of 473 nm and a zeta potential of –26 mV . The adsorption and elution of IgG on these adsorbents was thoroughly studied. Adsorption of human IgG was enhanced at pH 6, for which a q max of 244 mg IgG g −1 MPs and K d of 25 mg L −1 were obtained. Increasing salt concentrations at a basic pH (1 mol L −1 NaCl at pH 11) were found to improve the elution of bound IgG . The MPs were challenged with an artificial protein mixture containing human IgG , albumin, insulin and apo‐transferrin. An overall yield of 84% was achieved, retrieving 92% of bound IgG . CONCLUSIONS MPs were successfully used for the capture of monoclonal antibodies from two distinct mammalian cell cultures, a Chinese hamster ovary ( CHO ) and a hybridoma cell culture supernatants. The elution yields were high, ranging between 84% and 94%, with overall yields ranging from 72% to 88%. Final purities of 85% were reached for hybridoma cell supernatants. © 2014 Society of Chemical Industry