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dc.contributor.authorKorn, Janin
dc.contributor.authorSchäckermann, Dorina
dc.contributor.authorKirmann, Toni
dc.contributor.authorBertoglio, Federico
dc.contributor.authorSteinke, Stephan
dc.contributor.authorHeisig, Janyn
dc.contributor.authorRuschig, Maximilian
dc.contributor.authorRojas, Gertrudis
dc.contributor.authorLangreder, Nora
dc.contributor.authorWenzel, Esther Veronika
dc.contributor.authorRoth, Kristian Daniel Ralph
dc.contributor.authorBecker, Marlies
dc.contributor.authorMeier, Doris
dc.contributor.authorvan den Heuvel, Joop
dc.contributor.authorHust, Michael
dc.contributor.authorDübel, Stefan
dc.contributor.authorSchubert, Maren
dc.date.accessioned2021-01-04T13:43:47Z
dc.date.available2021-01-04T13:43:47Z
dc.date.issued2020-12-07
dc.identifier.citationSci Rep. 2020 Dec 7;10(1):21393. doi: 10.1038/s41598-020-78425-9.en_US
dc.identifier.pmid33288836
dc.identifier.doi10.1038/s41598-020-78425-9
dc.identifier.urihttp://hdl.handle.net/10033/622658
dc.description.abstractMammalian cells are the most commonly used production system for therapeutic antibodies. Protocols for the expression of recombinant antibodies in HEK293-6E cells in different antibody formats are described in detail. As model, antibodies against Kallikrein-related peptidase 7 (KLK7) were used. KLK7 is a key player in skin homeostasis and represents an emerging target for pharmacological interventions. Potent inhibitors can not only help to elucidate physiological and pathophysiological functions but also serve as a new archetype for the treatment of inflammatory skin disorders. Phage display-derived affinity-matured human anti-KLK7 antibodies were converted to scFv-Fc, IgG, and Fab formats and transiently produced in the mammalian HEK293-6E system. For the production of the corresponding antigen-KLK7-the baculovirus expression vector system (BEVS) and virus-free expression in Hi5 insect cells were used in a comparative approach. The target proteins were isolated by various chromatographic methods in a one- or multistep purification strategy. Ultimately, the interaction between anti-KLK7 and KLK7 was characterized using biolayer interferometry. Here, protocols for the expression of recombinant antibodies in different formats are presented and compared for their specific features. Furthermore, biolayer interferometry (BLI), a fast and high-throughput biophysical analytical technique to evaluate the kinetic binding constant and affinity constant of the different anti-KLK7 antibody formats against Kallikrein-related peptidase 7 is presented.en_US
dc.language.isoenen_US
dc.publisherNature researchen_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.titleBaculovirus-free insect cell expression system for high yield antibody and antigen production.en_US
dc.typeArticleen_US
dc.typeOtheren_US
dc.identifier.eissn2045-2322
dc.contributor.departmentHZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.en_US
dc.identifier.journalScientific reportsen_US
dc.source.volume10
dc.source.issue1
dc.source.beginpage21393
dc.source.endpage
refterms.dateFOA2021-01-04T13:43:48Z
dc.source.journaltitleScientific reports
dc.source.countryEngland


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Attribution 4.0 International
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