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dc.contributor.author
Pogson, Mark
dc.contributor.author
Parola, Cristina
dc.contributor.author
Kelton, William J.
dc.contributor.author
Heuberger, Paul
dc.contributor.author
Reddy, Sai T.
dc.date.accessioned
2018-09-12T06:14:42Z
dc.date.available
2018-09-12T06:14:42Z
dc.date.issued
2016
dc.identifier.issn
2041-1723
dc.identifier.other
10.1038/ncomms12535
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/288673
dc.identifier.doi
10.3929/ethz-b-000119720
dc.description.abstract
Hybridomas, fusions of primary mouse B cells and myelomas, are stable, rapidly-proliferating cell lines widely utilized for antibody screening and production. Antibody specificity of a hybridoma clone is determined by the immunoglobulin sequence of the primary B cell. Here we report a platform for rapid reprogramming of hybridoma antibody specificity by immunogenomic engineering. Here we use CRISPR-Cas9 to generate double-stranded breaks in immunoglobulin loci, enabling deletion of the native variable light chain and replacement of the endogenous variable heavy chain with a fluorescent reporter protein (mRuby). New antibody genes are introduced by Cas9-targeting of mRuby for replacement with a donor construct encoding a light chain and a variable heavy chain, resulting in full-length antibody expression. Since hybridomas surface express and secrete antibodies, reprogrammed cells are isolated using flow cytometry and cell culture supernatant is used for antibody production. Plug-and-(dis)play hybridomas can be reprogrammed with only a single transfection and screening step.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
Nature
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
Immunogenomic engineering of a plug-and-(dis)play hybridoma platform
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
dc.date.published
2016-08-17
ethz.journal.title
Nature Communications
ethz.journal.volume
7
en_US
ethz.journal.abbreviated
Nat Commun
ethz.pages.start
12535
en_US
ethz.size
10 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
London
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02060 - Dep. Biosysteme / Dep. of Biosystems Science and Eng.::03952 - Reddy, Sai / Reddy, Sai
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02060 - Dep. Biosysteme / Dep. of Biosystems Science and Eng.::03952 - Reddy, Sai / Reddy, Sai
ethz.date.deposited
2017-06-12T11:25:40Z
ethz.source
ECIT
ethz.identifier.importid
imp593654a6e913093882
ethz.identifier.importid
imp5936552f70a0967893
ethz.ecitpid
pub:181746
ethz.ecitpid
pub:190253
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2018-09-12T06:14:47Z
ethz.rosetta.lastUpdated
2024-02-02T06:02:45Z
ethz.rosetta.versionExported
true
dc.identifier.olduri
http://hdl.handle.net/20.500.11850/119720
dc.identifier.olduri
http://hdl.handle.net/20.500.11850/164661
ethz.COinS
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