title: Fishing human monoclonal antibodies from a CHO cell supernatant with boronic acid magnetic particles.
abstract: In this work we have evaluated the potential of boronic acid functionalized magnetic particles for the one-step capture of a human monoclonal antibody (mAb) from a Chinese hamster ovary (CHO) cell culture supernatant. For comparison, Protein A coated magnetic particles were also used. The most important factor influencing the overall process yield and product purity in boronic acid particles was found to be the binding pH. Basic pH values promoted higher purities while resulting in decreased yields due to the competing effects of molecules such as glucose and lactate present in the cell culture supernatant. After optimization, the particles were successfully used in a multi-cycle purification process of the mAb from the CHO feedstock. Boronic acid particles were able to achieve an average overall yield of 86% with 88% removal of CHO host cell proteins (HCP) when the binding was performed at pH 7.4, while at pH 8.5 these values were 58% and 97%, respectively. In both cases, genomic DNA removal was in excess of 97%. Comparatively, Protein A particles recorded an average overall yield of 80% and an HCP removal greater than 99%. The adsorption of the mAb to the boronic acid particles was shown to be mediated by strong affinity interactions. Overall, boronic acid based purification processes can offer a cost-effective alternative to Protein A as the direct capturing step from the mammalian cell culture.
authors.affiliation: Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Department of Bioengineering, Instituto Superior Técnico, Technical University of Lisbon, Av. Rovisco Pais, 1049-001 Lisbon, Portugal.
authors.forename: L
authors.initials: L
authors.lastname: Borlido
authors.forename: A M
authors.initials: AM
authors.lastname: Azevedo
authors.forename: A G
authors.initials: AG
authors.lastname: Sousa
authors.forename: P H
authors.initials: PH
authors.lastname: Oliveira
authors.forename: A C A
authors.initials: AC
authors.lastname: Roque
authors.forename: M R
authors.initials: MR
authors.lastname: Aires-Barros
mesh_terms: D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001897:Boronic Acids / Q000737:chemistry* / Q000378:metabolism; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D059346:Magnets / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D001897:Boronic Acids