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dc.contributor.author
Schwaigerlehner, Linda
dc.contributor.author
Pechlaner, Maria
dc.contributor.author
Mayrhofer, Patrick
dc.contributor.author
Oostenbrink, Chris
dc.contributor.author
Kunert, Renate
dc.date.accessioned
2019-01-18T13:11:58Z
dc.date.available
2018-12-22T10:37:14Z
dc.date.available
2019-01-18T13:11:58Z
dc.date.issued
2018
dc.identifier.issn
1741-0126
dc.identifier.issn
1741-0134
dc.identifier.other
10.1093/protein/gzy009
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/313168
dc.identifier.doi
10.3929/ethz-b-000313168
dc.description.abstract
Humanized monoclonal antibodies (mAbs) are among the most promising modern therapeutics, but defined engineering strategies are still not available. Antibody humanization often leads to a loss of affinity, as it is the case for our model antibody Ab2/3H6 (PDB entry 3BQU). Identifying appropriate back-to-mouse mutations is needed to restore binding affinity, but highly challenging. In order to get more insight, we have applied molecular dynamics simulations and correlated them to antibody binding and expression in wet lab experiments. In this study, we discuss six mAb variants and investigate a tyrosine conglomeration, an isopolar substitution and the improvement of antibody binding towards wildtype affinity. In the 3D structure of the mouse wildtype, residue R94h is surrounded by three tyrosines which form a so-called ‘tyrosine cage’. We demonstrate that the tyrosine cage has a supporting function for the CDRh3 loop conformation. The isopolar substitution is not able to mimic the function appropriately. Finally, we show that additional light chain mutations can restore binding to wildtype-comparable level, and also improve the expression of the mAb significantly. We conclude that the variable light chain of Ab2/3H6 is of underestimated importance for the interaction with its antigen mAb 2F5.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Oxford University Press
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.subject
antibody humanization
en_US
dc.subject
binding affinity
en_US
dc.subject
conformational clustering
en_US
dc.subject
GROMOS
en_US
dc.subject
molecular dynamics
en_US
dc.title
Lessons learned from merging wet lab experiments with molecular simulation to improve mAb humanization
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
dc.date.published
2018-05-11
ethz.journal.title
Protein Engineering, Design & Selection
ethz.journal.volume
31
en_US
ethz.journal.issue
7-8
en_US
ethz.journal.abbreviated
Protein Eng Des Sel
ethz.pages.start
257
en_US
ethz.pages.end
265
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
Oxford
en_US
ethz.publication.status
published
en_US
ethz.date.deposited
2018-12-22T10:37:16Z
ethz.source
WOS
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2019-01-18T13:12:08Z
ethz.rosetta.lastUpdated
2024-02-02T06:59:47Z
ethz.rosetta.versionExported
true
ethz.COinS
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