title: Optimizing expression and purification from cell culture medium of trispecific recombinant antibody derivatives.
abstract: Antibody fragments offer the possibility to build multifunctional manifolds tailored to meet a large variety of needs. We optimized the production of a manifold consisting of one (bibody) or two (tribody) single-chain variable fragments coupled to the C-terminus of Fab chains. Different strong mammalian promoters were compared and the influence of expression media on production and recovery was investigated. Since the physical and chemical nature of these molecules largely depends on the nature of the antibody building blocks incorporated, a generally applicable process for the purification of recombinant antibody derivatives from serum containing mammalian cell culture medium was designed. To this end we compared protein L, hydroxyapatite, immobilized metal affinity chromatography, cation-exchange chromatography and hydrophobic charge induction chromatography.
authors.affiliation: Department of Molecular Biomedical Research, Flanders Interuniversity Institute for Biotechnology (VIB), University of Ghent, K.L. Ledeganckstraat 35, 9000, Ghent, Belgium.
authors.forename: An
authors.initials: A
authors.lastname: Willems
authors.forename: Jannick
authors.initials: J
authors.lastname: Leoen
authors.forename: Steve
authors.initials: S
authors.lastname: Schoonooghe
authors.forename: Johan
authors.initials: J
authors.lastname: Grooten
authors.forename: Nico
authors.initials: N
authors.lastname: Mertens
mesh_terms: D002460:Cell Line; D002853:Chromatography, Liquid; D003470:Culture Media / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D003470:Culture Media; D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins