Glycan variability on a recombinant IgG antibody transiently produced in HEK-293E cells
| Autor: | Simone Schmitt, Yury O. Tsybin, Sophie Nallet, Lucia Baldi, Julien Parra, Luca Fornelli, Florian M. Wurm |
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| Jazyk: | angličtina |
| Předmět: |
Glycan
Glycosylation Gene-Expression Bioengineering Therapeutics Biology Transfection law.invention chemistry.chemical_compound Polysaccharides law Gene expression Suspension Protein biosynthesis Humans Amino Acid Sequence Molecular Biology Chromatography High Pressure Liquid HEK 293 cells Glycopeptides General Medicine Molecular biology Recombinant Proteins Mammalian-Cells HEK293 Cells chemistry Immunoglobulin G Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Recombinant DNA Antibody Protein-Production Biotechnology |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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