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Llama-Derived Single Domain Antibodies to Build Multivalent, Superpotent and Broadened Neutralizing Anti-Viral Molecules

Hultberg, A; Temperton, NJ; Rosseels, V; Koenders, M; Gonzalez-Pajuelo, M; Schepens, B; Ibanez, LI; ... de Haard, HJ; + view all (2011) Llama-Derived Single Domain Antibodies to Build Multivalent, Superpotent and Broadened Neutralizing Anti-Viral Molecules. PLOS ONE , 6 (4) , Article e17665. 10.1371/journal.pone.0017665. Green open access

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Abstract

For efficient prevention of viral infections and cross protection, simultaneous targeting of multiple viral epitopes is a powerful strategy. Llama heavy chain antibody fragments (VHH) against the trimeric envelope proteins of Respiratory Syncytial Virus (Fusion protein), Rabies virus (Glycoprotein) and H5N1 Influenza (Hemagglutinin 5) were selected from llama derived immune libraries by phage display. Neutralizing VHH recognizing different epitopes in the receptor binding sites on the spikes with affinities in the low nanomolar range were identified for all the three viruses by viral neutralization assays. By fusion of VHH with variable linker lengths, multimeric constructs were made that improved neutralization potencies up to 4,000-fold for RSV, 1,500-fold for Rabies virus and 75-fold for Influenza H5N1. The potencies of the VHH constructs were similar or better than best performing monoclonal antibodies. The cross protection capacity against different viral strains was also improved for all three viruses, both by multivalent (two or three identical VHH) and biparatopic (two different VHH) constructs. By combining a VHH neutralizing RSV subtype A, but not subtype B with a poorly neutralizing VHH with high affinity for subtype B, a biparatopic construct was made with low nanomolar neutralizing potency against both subtypes. Trivalent anti-H5N1 VHH neutralized both Influenza H5N1 clade1 and 2 in a pseudotype assay and was very potent in neutralizing the NIBRG-14 Influenza H5N1 strain with IC50 of 9 picomolar. Bivalent and biparatopic constructs against Rabies virus cross neutralized both 10 different Genotype 1 strains and Genotype 5. The results show that multimerization of VHH fragments targeting multiple epitopes on a viral trimeric spike protein is a powerful tool for anti-viral therapy to achieve "best-in-class" and broader neutralization capacity.

Type: Article
Title: Llama-Derived Single Domain Antibodies to Build Multivalent, Superpotent and Broadened Neutralizing Anti-Viral Molecules
Open access status: An open access version is available from UCL Discovery
DOI: 10.1371/journal.pone.0017665
Publisher version: http://dx.doi.org/10.1371/journal.pone.0017665
Language: English
Additional information: © 2011 Hultberg et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding was provided by an IWT grant (Institute for the Promotion of Innovation by Science and Technology in Flanders, IWT70050) from the Flemish government. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Funding was also provided by Ablynx NV which had a significant input on the design and execution of the research.
Keywords: RESPIRATORY SYNCYTIAL VIRUS, MONOCLONAL-ANTIBODY, RABIES VIRUS, IN-VITRO, ANTIGENIC STRUCTURE, F-GLYCOPROTEIN, ESCAPE MUTANTS, FRAGMENTS, MICE, VARIANTS
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Infection and Immunity
URI: https://discovery.ucl.ac.uk/id/eprint/1329778
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