Please use this identifier to cite or link to this item: https://doi.org/10.1038/s42003-021-01965-x
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dc.titleEpitope-directed monoclonal antibody production using a mixed antigen cocktail facilitates antibody characterization and validation
dc.contributor.authorLiew, Oi Wah
dc.contributor.authorLing, Samantha SM
dc.contributor.authorLilyanna, Shera
dc.contributor.authorZhou, Yue
dc.contributor.authorWang, Peipei
dc.contributor.authorChong, Jenny PC
dc.contributor.authorYan, Xia Ng
dc.contributor.authorLim, Angeline ES
dc.contributor.authorLeong, Eliot RY
dc.contributor.authorLin, Qifeng
dc.contributor.authorLim, Teck Kwang
dc.contributor.authorLin, Qingsong
dc.contributor.authorNg, Enoch MW
dc.contributor.authorNg, Tuck Wah
dc.contributor.authorRichards, A Mark
dc.date.accessioned2021-11-17T03:51:11Z
dc.date.available2021-11-17T03:51:11Z
dc.date.issued2021-04-06
dc.identifier.citationLiew, Oi Wah, Ling, Samantha SM, Lilyanna, Shera, Zhou, Yue, Wang, Peipei, Chong, Jenny PC, Yan, Xia Ng, Lim, Angeline ES, Leong, Eliot RY, Lin, Qifeng, Lim, Teck Kwang, Lin, Qingsong, Ng, Enoch MW, Ng, Tuck Wah, Richards, A Mark (2021-04-06). Epitope-directed monoclonal antibody production using a mixed antigen cocktail facilitates antibody characterization and validation. COMMUNICATIONS BIOLOGY 4 (1). ScholarBank@NUS Repository. https://doi.org/10.1038/s42003-021-01965-x
dc.identifier.issn23993642
dc.identifier.urihttps://scholarbank.nus.edu.sg/handle/10635/206550
dc.description.abstractHigh quality, well-validated antibodies are needed to mitigate irreproducibility and clarify conflicting data in science. We describe an epitope-directed monoclonal antibody (mAb) production method that addresses issues of antibody quality, validation and utility. The workflow is illustrated by generating mAbs against multiple in silico-predicted epitopes on human ankyrin repeat domain 1 (hANKRD1) in a single hybridoma production cycle. Antigenic peptides (13–24 residues long) presented as three-copy inserts on the surface exposed loop of a thioredoxin carrier produced high affinity mAbs that are reactive to native and denatured hANKRD1. ELISA assay miniaturization afforded by novel DEXT microplates allowed rapid hybridoma screening with concomitant epitope identification. Antibodies against spatially distant sites on hANKRD1 facilitated validation schemes applicable to two-site ELISA, western blotting and immunocytochemistry. The use of short antigenic peptides of known sequence facilitated direct epitope mapping crucial for antibody characterization. This robust method motivates its ready adoption for other protein targets.
dc.language.isoen
dc.publisherNATURE RESEARCH
dc.sourceElements
dc.subjectScience & Technology
dc.subjectLife Sciences & Biomedicine
dc.subjectBiology
dc.subjectMultidisciplinary Sciences
dc.subjectLife Sciences & Biomedicine - Other Topics
dc.subjectScience & Technology - Other Topics
dc.typeArticle
dc.date.updated2021-11-11T00:52:26Z
dc.contributor.departmentDEPT OF BIOLOGICAL SCIENCES
dc.contributor.departmentDEPT OF MEDICINE
dc.description.doi10.1038/s42003-021-01965-x
dc.description.sourcetitleCOMMUNICATIONS BIOLOGY
dc.description.volume4
dc.description.issue1
dc.published.statePublished
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