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dc.contributor.author
Schanz, Merle
dc.contributor.author
Liechti, Thomas
dc.contributor.author
Zagordi, Osvaldo
dc.contributor.author
Miho, Enkelejda
dc.contributor.author
Reddy, Sai T.
dc.contributor.author
Günthard, Huldrych F.
dc.contributor.author
Trkola, Alexandra
dc.contributor.author
Huber, Michael
dc.date.accessioned
2018-08-09T07:40:47Z
dc.date.available
2017-06-11T14:56:49Z
dc.date.available
2018-08-09T07:40:47Z
dc.date.issued
2014-11-03
dc.identifier.issn
1932-6203
dc.identifier.other
10.1371/journal.pone.0111726
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/95131
dc.identifier.doi
10.3929/ethz-b-000095131
dc.description.abstract
The humoral immune response plays a critical role in controlling infection, and the rapid adaptation to a broad range of pathogens depends on a highly diverse antibody repertoire. The advent of high-throughput sequencing technologies in the past decade has enabled insights into this immense diversity. However, not only the variable, but also the constant region of antibodies determines their in vivo activity. Antibody isotypes differ in effector functions and are thought to play a defining role in elicitation of immune responses, both in natural infection and in vaccination. We have developed an Illumina MiSeq high-throughput sequencing protocol that allows determination of the human IgG subtype alongside sequencing full-length antibody variable heavy chain regions. We thereby took advantage of the Illumina procedure containing two additional short reads as identifiers. By performing paired-end sequencing of the variable regions and customizing one of the identifier sequences to distinguish IgG subtypes, IgG transcripts with linked information of variable regions and IgG subtype can be retrieved. We applied our new method to the analysis of the IgG variable region repertoire from PBMC of an HIV-1 infected individual confirmed to have serum antibody reactivity to the Membrane Proximal External Region (MPER) of gp41. We found that IgG3 subtype frequencies in the memory B cell compartment increased after halted treatment and coincided with increased plasma antibody reactivity against the MPER domain. The sequencing strategy we developed is not restricted to analysis of IgG. It can be adopted for any Ig subtyping and beyond that for any research question where phasing of distant regions on the same amplicon is needed.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
PLOS
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.title
High-Throughput Sequencing of Human Immunoglobulin Variable Regions with Subtype Identification
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
ethz.journal.title
PLoS ONE
ethz.journal.volume
9
en_US
ethz.journal.issue
11
en_US
ethz.journal.abbreviated
PLoS ONE
ethz.pages.start
e111726
en_US
ethz.size
9 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.publication.place
San Francisco, CA
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
2017-06-11T14:56:53Z
ethz.source
ECIT
ethz.identifier.importid
imp593652b8f3d8513637
ethz.ecitpid
pub:149306
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-07-14T18:43:45Z
ethz.rosetta.lastUpdated
2024-02-02T05:37:49Z
ethz.rosetta.versionExported
true
ethz.COinS
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