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Llama-derived single domain antibodies to build multivalent, superpotent and broadened neutralizing anti-viral molecules

Anna Hultberg, Nigel J Temperton, Valérie Rosseels, Mireille Koenders, Maria Gonzalez-Pajuelo, Bert Schepens UGent, Lorena Ibanez UGent, Peter Vanlandschoot, Joris Schillemans and Michael Saunders, et al. (2011) PLOS ONE. 6(4).
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.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
MONOCLONAL-ANTIBODY, RESPIRATORY SYNCYTIAL VIRUS, VARIANTS, MICE, FRAGMENTS, ESCAPE MUTANTS, F-GLYCOPROTEIN, RABIES VIRUS, ANTIGENIC STRUCTURE, IN-VITRO
journal title
PLOS ONE
PLoS One
volume
6
issue
4
article_number
e17665
pages
12 pages
Web of Science type
Article
Web of Science id
000289058300003
JCR category
BIOLOGY
JCR impact factor
4.092 (2011)
JCR rank
12/84 (2011)
JCR quartile
1 (2011)
ISSN
1932-6203
DOI
10.1371/journal.pone.0017665
language
English
UGent publication?
yes
classification
A1
copyright statement
I have retained and own the full copyright for this publication
id
1212300
handle
http://hdl.handle.net/1854/LU-1212300
date created
2011-04-26 13:47:26
date last changed
2012-06-26 14:32:17
@article{1212300,
  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 {\textacutedbl}best-in-class{\textacutedbl} and broader neutralization capacity.},
  articleno    = {e17665},
  author       = {Hultberg, Anna and Temperton, Nigel J and Rosseels, Val{\'e}rie and Koenders, Mireille and Gonzalez-Pajuelo, Maria and Schepens, Bert and Ibanez, Lorena and Vanlandschoot, Peter and Schillemans, Joris and Saunders, Michael and Weiss, Robin A and Saelens, Xavier and Melero, Jos{\'e} A and Verrips, C Theo and Van Gucht, Steven and de Haard, Hans J},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keyword      = {MONOCLONAL-ANTIBODY,RESPIRATORY SYNCYTIAL VIRUS,VARIANTS,MICE,FRAGMENTS,ESCAPE MUTANTS,F-GLYCOPROTEIN,RABIES VIRUS,ANTIGENIC STRUCTURE,IN-VITRO},
  language     = {eng},
  number       = {4},
  pages        = {12},
  title        = {Llama-derived single domain antibodies to build multivalent, superpotent and broadened neutralizing anti-viral molecules},
  url          = {http://dx.doi.org/10.1371/journal.pone.0017665},
  volume       = {6},
  year         = {2011},
}

Chicago
Hultberg, Anna, Nigel J Temperton, Valérie Rosseels, Mireille Koenders, Maria Gonzalez-Pajuelo, Bert Schepens, Lorena Ibanez, et al. 2011. “Llama-derived Single Domain Antibodies to Build Multivalent, Superpotent and Broadened Neutralizing Anti-viral Molecules.” Plos One 6 (4).
APA
Hultberg, A., Temperton, N. J., Rosseels, V., Koenders, M., Gonzalez-Pajuelo, M., Schepens, B., Ibanez, L., et al. (2011). Llama-derived single domain antibodies to build multivalent, superpotent and broadened neutralizing anti-viral molecules. PLOS ONE, 6(4).
Vancouver
1.
Hultberg A, Temperton NJ, Rosseels V, Koenders M, Gonzalez-Pajuelo M, Schepens B, et al. Llama-derived single domain antibodies to build multivalent, superpotent and broadened neutralizing anti-viral molecules. PLOS ONE. 2011;6(4).
MLA
Hultberg, Anna, Nigel J Temperton, Valérie Rosseels, et al. “Llama-derived Single Domain Antibodies to Build Multivalent, Superpotent and Broadened Neutralizing Anti-viral Molecules.” PLOS ONE 6.4 (2011): n. pag. Print.