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Wagner, Ralf ; Bruun, Tim-Henrik ; Mühlbauer, Katharina ; Benen, Thomas ; Kliche, Alexander

A mammalian cell based FACS-panning platform for the selection of HIV-1 envelopes for vaccine development

Wagner, Ralf, Bruun, Tim-Henrik, Mühlbauer, Katharina, Benen, Thomas and Kliche, Alexander (2014) A mammalian cell based FACS-panning platform for the selection of HIV-1 envelopes for vaccine development. PLoS ONE 9 (10), e109196.

Date of publication of this fulltext: 10 Oct 2014 11:07
Article
DOI to cite this document: 10.5283/epub.30861


Abstract

An increasing number of broadly neutralizing monoclonal antibodies (bnMAb) against the HIV-1 envelope (Env) protein has been discovered recently. Despite this progress, vaccination efforts with the aim to re-elicit bnMAbs that provide protective immunity have failed so far. Herein, we describe the development of a mammalian cell based FACS-panning method in which bnMAbs are used as tools to ...

An increasing number of broadly neutralizing monoclonal antibodies (bnMAb) against the HIV-1 envelope (Env) protein has been discovered recently. Despite this progress, vaccination efforts with the aim to re-elicit bnMAbs that provide protective immunity have failed so far. Herein, we describe the development of a mammalian cell based FACS-panning method in which bnMAbs are used as tools to select surface-exposed envelope variants according to their binding affinity. For that purpose, an HIV-1 derived lentiviral vector was developed to infect HEK293T cells at low multiplicity of infection (MOI) in order to link Env phenotype and genotype. For proof of principle, a gp145 Env model-library was established in which the complete V3 domain was substituted by five strain specific V3 loop sequences with known binding affinities to nMAb 447-52D, respectively. Env genes were recovered from selected cells by PCR, subcloned into a lentiviral vector (i) to determine and quantify the enrichment nMAb binders and (ii) to generate a new batch of transduction competent particles. After 2 selection cycles the Env variant with highest affinity was enriched 20-fold and represented 80% of the remaining Env population. Exploiting the recently described bnMAbs, this procedure might prove useful in selecting Env proteins from large Env libraries with the potential to elicit bnMAbs when used as vaccine candidates.



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Details

Item typeArticle
Journal or Publication TitlePLoS ONE
Publisher:PUBLIC LIBRARY SCIENCE
Place of Publication:SAN FRANCISCO
Volume:9
Number of Issue or Book Chapter:10
Page Range:e109196
Date3 October 2014
InstitutionsMedicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene
Identification Number
ValueType
10.1371/journal.pone.0109196DOI
KeywordsIMMUNODEFICIENCY-VIRUS TYPE-1; HUMAN MONOCLONAL-ANTIBODIES; TIME QUANTITATIVE PCR; NEUTRALIZING ANTIBODIES; IN-VIVO; ONE-POT; DESIGN; GENE; DISPLAY; VECTOR;
Dewey Decimal Classification600 Technology > 610 Medical sciences Medicine
StatusPublished
RefereedYes, this version has been refereed
Created at the University of RegensburgYes
URN of the UB Regensburgurn:nbn:de:bvb:355-epub-308615
Item ID30861

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