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dc.contributor.authorTaube, Ran
dc.contributor.authorZhu, Quan Karen
dc.contributor.authorXu, Chen
dc.contributor.authorDiaz-Griffero, Felipe
dc.contributor.authorSui, Jianhua
dc.contributor.authorKamau, Erick
dc.contributor.authorDwyer, Markryan
dc.contributor.authorAird, Daniel
dc.contributor.authorMarasco, Wayne A.
dc.date.accessioned2011-02-12T04:16:51Z
dc.date.issued2008
dc.identifier.citationTaube, Ran, Quan Zhu, Chen Xu, Felipe Diaz-Griffero, Jianhua Sui, Erick Kamau, Markryan Dwyer, Daniel Aird, and Wayne A. Marasco. 2008. Lentivirus display: Stable expression of human antibodies on the surface of human cells and virus particles. PLoS ONE 3(9): e3181.en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:4724768
dc.description.abstractBackground: Isolation of human antibodies using current display technologies can be limited by constraints on protein expression, folding and post-translational modifications. Here we describe a discovery platform that utilizes self-inactivating (SIN) lentiviral vectors for the surface display of high-affinity single-chain variable region (scFv) antibody fragments on human cells and lentivirus particles. Methodology/Principal Findings: Bivalent scFvFc human antibodies were fused in frame with different transmembrane (TM) anchoring moieties to allow efficient high-level expression on human cells and the optimal TM was identified. The addition of an eight amino acid HIV-1 gp41 envelope incorporation motif further increased scFvFc expression on human cells and incorporation into lentiviral particles. Both antibody-displaying human cells and virus particles bound antigen specifically. Sulfation of CDR tyrosine residues, a property recently shown to broaden antibody binding affinity and antigen recognition was also demonstrated. High level scFvFc expression and stable integration was achieved in human cells following transduction with IRES containing bicistronic SIN lentivectors encoding ZsGreen when scFvFc fusion proteins were expressed from the first cassette. Up to 10^6-fold enrichment of antibody expressing cells was achieved with one round of antigen coupled magnetic bead pre-selection followed by FACS sorting. Finally, the scFvFc displaying human cells could be used directly in functional biological screens with remarkable sensitivity. Conclusions/Significance: This antibody display platform will complement existing technologies by virtue of providing properties unique to lentiviruses and antibody expression in human cells, which, in turn, may aid the discovery of novel therapeutic human mAbs.en_US
dc.language.isoen_USen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofdoi://10.1371/journal.pone.0003181en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2527531/pdf/en_US
dash.licenseLAA
dc.subjectImmunologyen_US
dc.subjectMolecular Biologyen_US
dc.subjectBiotechnology/Bioengineeringen_US
dc.titleLentivirus Display: Stable Expression of Human Antibodies on the Surface of Human Cells and Virus Particlesen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalPLoS ONEen_US
dash.depositing.authorMarasco, Wayne A.
dc.date.available2011-02-12T04:16:51Z
dash.affiliation.otherHMS^Medicine-Brigham and Women's Hospitalen_US
dash.affiliation.otherHMS^Medicine-Brigham and Women's Hospitalen_US
dc.identifier.doi10.1371/journal.pone.0003181*
dash.contributor.affiliatedMarasco, Wayne
dash.contributor.affiliatedZhu, Quan


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