title: Insect cells for antibody production: evaluation of an efficient alternative.
abstract: In recent years there has been an increase in both availability and demand for therapeutic monoclonal antibodies. Currently, most of these antibodies are produced by stably transfected mammalian cells. In this study we evaluated the use of different baculoviral insect cell systems as an alternative for commonly used production schemes. We expressed the human anti-gp41 antibody 3D6 in Spodoptera frugiperda Sf9, Trichoplusia ni BTI-TN5B1-4 "High Five", and Spodoptera frugiperda SfSWT-1 "Mimic™" insect cells and compared product yield, specificity and glycosylation patterns with a 3D6 antibody expressed in Chinese hamster ovary cells. Using "High Five" cells we achieved amounts of secreted antibody comparable to those resulting from transient expression in mammalian cells. We determined the N-linked oligosaccharide structures present on asparagine-297 in IgG₁ heavy chains and tested the functionality in terms of antigen binding and the ability to elicit effector functions. Antibodies expressed in all insect cell lines displayed highly specific antigen binding. In general, the insect-produced antibodies carried, as the CHO-produced form, fucosylated N-glycans, including, in the case of "High Five" cells, high levels of core α1,3-fucose. This indicates that in all systems glycoengineering may be required in order to produce optimal glycoforms of this antibody.
authors.affiliation: Vienna Institute of BioTechnology, University of Natural Resources and Life Sciences, Muthgasse 11, 1190 Vienna, Austria.
authors.forename: Dieter
authors.initials: D
authors.lastname: Palmberger
authors.forename: Dubravko
authors.initials: D
authors.lastname: Rendić
authors.forename: Philipp
authors.initials: P
authors.lastname: Tauber
authors.forename: Florian
authors.initials: F
authors.lastname: Krammer
authors.forename: Iain B H
authors.initials: IB
authors.lastname: Wilson
authors.forename: Reingard
authors.initials: R
authors.lastname: Grabherr
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005643:Fucose / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism* / Q000821:virology
chemical_list: D000911:Antibodies, Monoclonal; D005643:Fucose