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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 und 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.Veröffentlichungsdatum dieses Volltextes: 10 Okt 2014 11:07
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DOI zum Zitieren dieses Dokuments: 10.5283/epub.30861
Zusammenfassung
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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| Dokumentenart | Artikel | ||||
| Titel eines Journals oder einer Zeitschrift | PLoS ONE | ||||
| Verlag: | PUBLIC LIBRARY SCIENCE | ||||
|---|---|---|---|---|---|
| Ort der Veröffentlichung: | SAN FRANCISCO | ||||
| Band: | 9 | ||||
| Nummer des Zeitschriftenheftes oder des Kapitels: | 10 | ||||
| Seitenbereich: | e109196 | ||||
| Datum | 3 Oktober 2014 | ||||
| Institutionen | Medizin > Lehrstuhl für Medizinische Mikrobiologie und Hygiene | ||||
| Identifikationsnummer |
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| Stichwörter / Keywords | IMMUNODEFICIENCY-VIRUS TYPE-1; HUMAN MONOCLONAL-ANTIBODIES; TIME QUANTITATIVE PCR; NEUTRALIZING ANTIBODIES; IN-VIVO; ONE-POT; DESIGN; GENE; DISPLAY; VECTOR; | ||||
| Dewey-Dezimal-Klassifikation | 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin | ||||
| Status | Veröffentlicht | ||||
| Begutachtet | Ja, diese Version wurde begutachtet | ||||
| An der Universität Regensburg entstanden | Ja | ||||
| URN der UB Regensburg | urn:nbn:de:bvb:355-epub-308615 | ||||
| Dokumenten-ID | 30861 |
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