Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.

Previously, our group engineered a plant-derived monoclonal antibody (MAb) (pHu-E16) that efficiently treated West Nile virus (WNV) infection in mice. In this study, we developed several pHu-E16 variants to improve its efficacy. These variants included a single-chain variable fragment (scFv) of pHu-...

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Autores principales:	Junyun He, Huafang Lai, Michael Engle, Sergey Gorlatov, Clemens Gruber, Herta Steinkellner, Michael S Diamond, Qiang Chen
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/af8673017dc749acbda25f198198c2f4
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spelling	oai:doaj.org-article:af8673017dc749acbda25f198198c2f42021-11-18T08:25:50ZGeneration and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.1932-620310.1371/journal.pone.0093541https://doaj.org/article/af8673017dc749acbda25f198198c2f42014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24675995/?tool=EBIhttps://doaj.org/toc/1932-6203Previously, our group engineered a plant-derived monoclonal antibody (MAb) (pHu-E16) that efficiently treated West Nile virus (WNV) infection in mice. In this study, we developed several pHu-E16 variants to improve its efficacy. These variants included a single-chain variable fragment (scFv) of pHu-E16 fused to the heavy chain (HC) constant domains (CH(1-3)) of human IgG (pHu-E16scFv-CH(1-3)) and a tetravalent molecule (Tetra pHu-E16) assembled from pHu-E16scFv-CH(1-3) with a second pHu-E16scFv fused to the light chain (LC) constant region. pHu-E16scFv-CH(1-3) and Tetra pHu-E16 were efficiently expressed and assembled in plants. To assess the impact of differences in N-linked glycosylation on pHu-E16 variant assembly and function, we expressed additional pHu-E16 variants with various combinations of HC and LC components. Our study revealed that proper pairing of HC and LC was essential for the complete N-glycan processing of antibodies in both plant and animal cells. Associated with their distinct N-glycoforms, pHu-E16, pHu-E16scFv-CH(1-3) and Tetra pHu-E16 exhibited differential binding to C1q and specific Fcγ receptors (FcγR). Notably, none of the plant-derived Hu-E16 variants showed antibody-dependent enhancement (ADE) activity in CD32A+ human cells, suggesting the potential of plant-produced antibodies to minimize the adverse effect of ADE. Importantly, all plant-derived MAb variants exhibited at least equivalent in vitro neutralization and in vivo protection in mice compared to mammalian cell-produced Hu-E16. This study demonstrates the capacity of plants to express and assemble a large, complex and functional IgG-like tetravalent mAb variant and also provides insight into the relationship between MAb N-glycosylation, FcγR and C1q binding, and ADE. These new insights may allow the development of safer and cost effective MAb-based therapeutics for flaviviruses, and possibly other pathogens.Junyun HeHuafang LaiMichael EngleSergey GorlatovClemens GruberHerta SteinkellnerMichael S DiamondQiang ChenPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 3, p e93541 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Junyun He Huafang Lai Michael Engle Sergey Gorlatov Clemens Gruber Herta Steinkellner Michael S Diamond Qiang Chen Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.
description	Previously, our group engineered a plant-derived monoclonal antibody (MAb) (pHu-E16) that efficiently treated West Nile virus (WNV) infection in mice. In this study, we developed several pHu-E16 variants to improve its efficacy. These variants included a single-chain variable fragment (scFv) of pHu-E16 fused to the heavy chain (HC) constant domains (CH(1-3)) of human IgG (pHu-E16scFv-CH(1-3)) and a tetravalent molecule (Tetra pHu-E16) assembled from pHu-E16scFv-CH(1-3) with a second pHu-E16scFv fused to the light chain (LC) constant region. pHu-E16scFv-CH(1-3) and Tetra pHu-E16 were efficiently expressed and assembled in plants. To assess the impact of differences in N-linked glycosylation on pHu-E16 variant assembly and function, we expressed additional pHu-E16 variants with various combinations of HC and LC components. Our study revealed that proper pairing of HC and LC was essential for the complete N-glycan processing of antibodies in both plant and animal cells. Associated with their distinct N-glycoforms, pHu-E16, pHu-E16scFv-CH(1-3) and Tetra pHu-E16 exhibited differential binding to C1q and specific Fcγ receptors (FcγR). Notably, none of the plant-derived Hu-E16 variants showed antibody-dependent enhancement (ADE) activity in CD32A+ human cells, suggesting the potential of plant-produced antibodies to minimize the adverse effect of ADE. Importantly, all plant-derived MAb variants exhibited at least equivalent in vitro neutralization and in vivo protection in mice compared to mammalian cell-produced Hu-E16. This study demonstrates the capacity of plants to express and assemble a large, complex and functional IgG-like tetravalent mAb variant and also provides insight into the relationship between MAb N-glycosylation, FcγR and C1q binding, and ADE. These new insights may allow the development of safer and cost effective MAb-based therapeutics for flaviviruses, and possibly other pathogens.
format	article
author	Junyun He Huafang Lai Michael Engle Sergey Gorlatov Clemens Gruber Herta Steinkellner Michael S Diamond Qiang Chen
author_facet	Junyun He Huafang Lai Michael Engle Sergey Gorlatov Clemens Gruber Herta Steinkellner Michael S Diamond Qiang Chen
author_sort	Junyun He
title	Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.
title_short	Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.
title_full	Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.
title_fullStr	Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.
title_full_unstemmed	Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.
title_sort	generation and analysis of novel plant-derived antibody-based therapeutic molecules against west nile virus.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/af8673017dc749acbda25f198198c2f4
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Generation and analysis of novel plant-derived antibody-based therapeutic molecules against West Nile virus.

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