Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea

Noroviruses are a major cause of acute gastroenteritis, but no vaccines or therapeutic drugs are available. Llama-derived single chain antibody fragments (also called VHH) are small, recombinant monoclonal antibodies of 15 kDa with several advantages over conventional antibodies. The aim of this stu...

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Main Authors:	Garaicoechea, Lorena, Aguilar, Andrea, Parra, Gabriel I., Bok, Marina, Sosnovtsev, Stanislav V., Canziani, Gabriela, Green, Kim Y., Bok, Karin, Parreño, Viviana
Format:	Online
Language:	English
Published:	Public Library of Science 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4534396/

id	pubmed-4534396
recordtype	oai_dc
spelling	pubmed-45343962015-08-24 Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea Garaicoechea, Lorena Aguilar, Andrea Parra, Gabriel I. Bok, Marina Sosnovtsev, Stanislav V. Canziani, Gabriela Green, Kim Y. Bok, Karin Parreño, Viviana Research Article Noroviruses are a major cause of acute gastroenteritis, but no vaccines or therapeutic drugs are available. Llama-derived single chain antibody fragments (also called VHH) are small, recombinant monoclonal antibodies of 15 kDa with several advantages over conventional antibodies. The aim of this study was to generate recombinant monoclonal VHH specific for the two major norovirus (NoV) genogroups (GI and GII) in order to investigate their potential as immunotherapy for the treatment of NoV diarrhea. To accomplish this objective, two llamas were immunized with either GI.1 (Norwalk-1968) or GII.4 (MD2004) VLPs. After immunization, peripheral blood lymphocytes were collected and used to generate two VHH libraries. Using phage display technology, 10 VHH clones specific for GI.1, and 8 specific for GII.4 were selected for further characterization. All VHH recognized conformational epitopes in the P domain of the immunizing VP1 capsid protein, with the exception of one GII.4 VHH that recognized a linear P domain epitope. The GI.1 VHHs were highly specific for the immunizing GI.1 genotype, with only one VHH cross-reacting with GI.3 genotype. The GII.4 VHHs reacted with the immunizing GII.4 strain and showed a varying reactivity profile among different GII genotypes. One VHH specific for GI.1 and three specific for GII.4 could block the binding of homologous VLPs to synthetic HBGA carbohydrates, saliva, and pig gastric mucin, and in addition, could inhibit the hemagglutination of red blood cells by homologous VLPs. The ability of Nov-specific VHHs to perform well in these surrogate neutralization assays supports their further development as immunotherapy for NoV treatment and immunoprophylaxis. Public Library of Science 2015-08-12 /pmc/articles/PMC4534396/ /pubmed/26267898 http://dx.doi.org/10.1371/journal.pone.0133665 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Garaicoechea, Lorena Aguilar, Andrea Parra, Gabriel I. Bok, Marina Sosnovtsev, Stanislav V. Canziani, Gabriela Green, Kim Y. Bok, Karin Parreño, Viviana
spellingShingle	Garaicoechea, Lorena Aguilar, Andrea Parra, Gabriel I. Bok, Marina Sosnovtsev, Stanislav V. Canziani, Gabriela Green, Kim Y. Bok, Karin Parreño, Viviana Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea
author_facet	Garaicoechea, Lorena Aguilar, Andrea Parra, Gabriel I. Bok, Marina Sosnovtsev, Stanislav V. Canziani, Gabriela Green, Kim Y. Bok, Karin Parreño, Viviana
author_sort	Garaicoechea, Lorena
title	Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea
title_short	Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea
title_full	Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea
title_fullStr	Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea
title_full_unstemmed	Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea
title_sort	llama nanoantibodies with therapeutic potential against human norovirus diarrhea
description	Noroviruses are a major cause of acute gastroenteritis, but no vaccines or therapeutic drugs are available. Llama-derived single chain antibody fragments (also called VHH) are small, recombinant monoclonal antibodies of 15 kDa with several advantages over conventional antibodies. The aim of this study was to generate recombinant monoclonal VHH specific for the two major norovirus (NoV) genogroups (GI and GII) in order to investigate their potential as immunotherapy for the treatment of NoV diarrhea. To accomplish this objective, two llamas were immunized with either GI.1 (Norwalk-1968) or GII.4 (MD2004) VLPs. After immunization, peripheral blood lymphocytes were collected and used to generate two VHH libraries. Using phage display technology, 10 VHH clones specific for GI.1, and 8 specific for GII.4 were selected for further characterization. All VHH recognized conformational epitopes in the P domain of the immunizing VP1 capsid protein, with the exception of one GII.4 VHH that recognized a linear P domain epitope. The GI.1 VHHs were highly specific for the immunizing GI.1 genotype, with only one VHH cross-reacting with GI.3 genotype. The GII.4 VHHs reacted with the immunizing GII.4 strain and showed a varying reactivity profile among different GII genotypes. One VHH specific for GI.1 and three specific for GII.4 could block the binding of homologous VLPs to synthetic HBGA carbohydrates, saliva, and pig gastric mucin, and in addition, could inhibit the hemagglutination of red blood cells by homologous VLPs. The ability of Nov-specific VHHs to perform well in these surrogate neutralization assays supports their further development as immunotherapy for NoV treatment and immunoprophylaxis.
publisher	Public Library of Science
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4534396/
_version_	1613258873900105728

Llama Nanoantibodies with Therapeutic Potential against Human Norovirus Diarrhea

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