Glycan variability on a recombinant IgG antibody transiently produced in HEK-293E cells.

Autor: Nallet S; École Polytechnique Fédérale de Lausanne, School of Life Sciences, Laboratory of Cellular Biotechnology, Station 6, CH-1015 Lausanne, Switzerland., Fornelli L, Schmitt S, Parra J, Baldi L, Tsybin YO, Wurm FM
Jazyk: angličtina
Zdroj: New biotechnology [N Biotechnol] 2012 May 15; Vol. 29 (4), pp. 471-6. Date of Electronic Publication: 2012 Mar 06.
DOI: 10.1016/j.nbt.2012.02.003
Abstrakt: 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.
(Copyright © 2012 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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