Llama-derived single domain antibodies to build multivalent, superpotent and broadened neutralizing anti-viral molecules
Hultberg, Anna, Temperton, Nigel J., Rosseels, Valerie, Koenders, Mireille, Gonzalez-Pajuelo, Maria, Schepens, Bert, Ibanez, Lorena Itatı, Vanlandschoot, Peter, Schillemans, Joris, Saunders, Michael, Weiss, Robin A., Saelens, Xavier, Melero, Jose A., Verrips, C. Theo, 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 (on-line) (doi:10.1371/journal.pone.0017665)
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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 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.
|Additional Information:|| 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.  Citation: Hultberg A, Temperton NJ, Rosseels V, Koenders M, Gonzalez-Pajuelo M, et al. (2011) Llama-Derived Single Domain Antibodies to Build Multivalent, Superpotent and Broadened Neutralizing Anti-Viral Molecules. PLoS ONE 6(4): e17665. doi:10.1371/journal.pone.0017665  Data was used for a patent application WO2009/147248 for which AH, PV, HJdH, CTV, MS, SvG, JAM, RAW, NJT, XS, and BS are inventors|
|Uncontrolled Keywords:||viral epitope, Llama, antibody fragments, respiratory syncytial virus, rabies virus, H5N1 Influenza|
|Subjects:||R Medicine > RC Internal medicine
R Medicine > RZ Other systems of medicine
|Faculty / Department / Research Groups:||Faculty of Engineering & Science > Medway School of Pharmacy|
|Last Modified:||18 Nov 2016 13:34|
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