Hultberg, Anna and Temperton, Nigel J. and Rosseels, Valerie and Koenders, Mireille and Gonzalez-Pajuelo, Maria and Schepens, Bert and Ibañez,, Lorena Itatí and Vanlandschoot, Peter and Schillemans, Joris and Saunders, Michael and Weiss, Robin A. and Saelens, Xavier and Melero, Jose A. and Verrips, Theo and van Gucht, Steven and de Haard, Hans J. (2011) Llama-derived single domain antibodies to build multivalent, superpotent and broadened neutralizing anti-viral molecules. PLoS ONE, 6 (4). pp. 1-12. ISSN 1932-6203. (doi:10.1371/journal.pone.0017665) (Full text available)
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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 IC 50 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. Â© 2011 Hultberg et al.
|Uncontrolled keywords:||epitope, glycoprotein, heavy chain antibody fragment, hybrid protein, immunoglobulin heavy chain, influenza B12 heavy chain antibody fragment, influenza C8 heavy chain antibody fragment, neutralizing antibody, palivizumab, rabies C12 heavy chain antibody fragment, rabies E6 heavy chain antibody fragment, rabies E8 heavy chain antibody fragment, rabies F8 heavy chain antibody fragment, rabies H7 heavy chain antibody fragment, respiratory syncytial virus 17 heavy chain antibody fragment, respiratory syncytial virus C4 heavy chain antibody fragment, respiratory syncytial virus D3 heavy chain antibody fragment, respiratory syncytial virus E4 heavy chain antibody fragment, respiratory syncytial virus NR1 heavy chain antibody fragment, respiratory syncytial virus NR4 heavy chain antibody fragment, unclassified drug, animal cell, antiviral therapy, article, Artiodactyla, binding site, controlled study, cross protection, drug potency, drug protein binding, genotype, human, human cell, IC 50, in vitro study, Influenza virus A H5N1, llama, nonhuman, nucleotide sequence, phage display, pseudotyping, Rabies virus, receptor binding, Respiratory syncytial pneumovirus, virus neutralization, Lama (mammal), Rabies virus, Respiratory syncytial virus, Rice stripe virus|
|Subjects:||Q Science > QR Microbiology > QR355 Virology|
|Divisions:||Faculties > Sciences > Medway School of Pharmacy|
|Depositing User:||Nigel Temperton|
|Date Deposited:||02 Aug 2012 07:01 UTC|
|Last Modified:||18 Jul 2014 11:13 UTC|
|Resource URI:||https://kar.kent.ac.uk/id/eprint/29398 (The current URI for this page, for reference purposes)|
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