title: Glycan variability on a recombinant IgG antibody transiently produced in HEK-293E cells.
abstract: In this study, a recombinant monoclonal IgG antibody was produced by transient gene expression (TGE) in suspension-adapted HEK-293E cells. The objective of the study was to determine the variation in recombinant IgG yield and glycosylation in ten independent transfections. In a ten-day batch process, the variation in transient IgG yield in the ten batches was less than 30% with the specific productivity averaging 20.2 ± 2.6 pg/cell/day. We characterized the N-glycosylation profile of each batch of affinity-purified IgG by intact protein and bottom-up mass spectrometry. Four major glycans were identified at Asn(297) in the ten batches with the maximum relative deviation for a single glycoform being 2.5%. In addition, within any single transfection there was little variation in glycoforms over the ten-day culture. Our experimental data indicate that with TGE, the production of recombinant IgG with little batch-to-batch variation in volumetric yield and protein glycosylation is feasible, even in a non-instrumented cultivation system as described here.
authors.affiliation: École Polytechnique Fédérale de Lausanne, School of Life Sciences, Laboratory of Cellular Biotechnology, Station 6, CH-1015 Lausanne, Switzerland.
authors.forename: Sophie
authors.initials: S
authors.lastname: Nallet
authors.forename: Luca
authors.initials: L
authors.lastname: Fornelli
authors.forename: Simone
authors.initials: S
authors.lastname: Schmitt
authors.forename: Julien
authors.initials: J
authors.lastname: Parra
authors.forename: Lucia
authors.initials: L
authors.lastname: Baldi
authors.forename: Yury O
authors.initials: YO
authors.lastname: Tsybin
authors.forename: Florian M
authors.initials: FM
authors.lastname: Wurm
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D006020:Glycopeptides / Q000032:analysis; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D011134:Polysaccharides / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006020:Glycopeptides; D007074:Immunoglobulin G; D011134:Polysaccharides; D011994:Recombinant Proteins