Dramatic activation of an antibody by a single amino acid change in framework

Abstract Antibody function is typically entirely dictated by the Complementarity Determining Regions (CDRs) that directly bind to the antigen, while the framework region acts as a scaffold for the CDRs and maintains overall structure of the variable domain. We recently reported that the rabbit monoc...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Wei-Ching Liang, Jianping Yin, Patrick Lupardus, Jianhuan Zhang, Kelly M. Loyet, Jawahar Sudhamsu, Yan Wu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0b851df1980f44c9ad9fd78eb3510ab2
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:0b851df1980f44c9ad9fd78eb3510ab2
record_format	dspace
spelling	oai:doaj.org-article:0b851df1980f44c9ad9fd78eb3510ab22021-11-21T12:25:11ZDramatic activation of an antibody by a single amino acid change in framework10.1038/s41598-021-01530-w2045-2322https://doaj.org/article/0b851df1980f44c9ad9fd78eb3510ab22021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01530-whttps://doaj.org/toc/2045-2322Abstract Antibody function is typically entirely dictated by the Complementarity Determining Regions (CDRs) that directly bind to the antigen, while the framework region acts as a scaffold for the CDRs and maintains overall structure of the variable domain. We recently reported that the rabbit monoclonal antibody 4A11 (rbt4A11) disrupts signaling through both TGFβ2 and TGFβ3 (Sun et al. in Sci Transl Med, 2021. https://doi.org/10.1126/scitranslmed.abe0407 ). Here, we report a dramatic, unexpected discovery during the humanization of rbt4A11 where, two variants of humanized 4A11 (h4A11), v2 and v7 had identical CDRs, maintained high affinity binding to TGFβ2/3, yet exhibited distinct differences in activity. While h4A11.v7 completely inhibited TGFβ2/3 signaling like rbt4A11, h4A11.v2 did not. We solved crystal structures of TGFβ2 complexed with Fab fragments of h4A11.v2 or h4A11.v7 and identified a novel interaction between the two heavy chain molecules in the 2:2 TGFb2:h4A11.v2-Fab complex. Further characterization revealed that framework residue variations at either position 19, 79 or 81 (Kabat numbering) of the heavy chain strikingly converts h4A11.v2 into an inhibitory antibody. Our work suggests that in addition to CDRs, framework residues and interactions between Fabs in an antibody could be engineered to further modulate activity of antibodies.Wei-Ching LiangJianping YinPatrick LupardusJianhuan ZhangKelly M. LoyetJawahar SudhamsuYan WuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Wei-Ching Liang Jianping Yin Patrick Lupardus Jianhuan Zhang Kelly M. Loyet Jawahar Sudhamsu Yan Wu Dramatic activation of an antibody by a single amino acid change in framework
description	Abstract Antibody function is typically entirely dictated by the Complementarity Determining Regions (CDRs) that directly bind to the antigen, while the framework region acts as a scaffold for the CDRs and maintains overall structure of the variable domain. We recently reported that the rabbit monoclonal antibody 4A11 (rbt4A11) disrupts signaling through both TGFβ2 and TGFβ3 (Sun et al. in Sci Transl Med, 2021. https://doi.org/10.1126/scitranslmed.abe0407 ). Here, we report a dramatic, unexpected discovery during the humanization of rbt4A11 where, two variants of humanized 4A11 (h4A11), v2 and v7 had identical CDRs, maintained high affinity binding to TGFβ2/3, yet exhibited distinct differences in activity. While h4A11.v7 completely inhibited TGFβ2/3 signaling like rbt4A11, h4A11.v2 did not. We solved crystal structures of TGFβ2 complexed with Fab fragments of h4A11.v2 or h4A11.v7 and identified a novel interaction between the two heavy chain molecules in the 2:2 TGFb2:h4A11.v2-Fab complex. Further characterization revealed that framework residue variations at either position 19, 79 or 81 (Kabat numbering) of the heavy chain strikingly converts h4A11.v2 into an inhibitory antibody. Our work suggests that in addition to CDRs, framework residues and interactions between Fabs in an antibody could be engineered to further modulate activity of antibodies.
format	article
author	Wei-Ching Liang Jianping Yin Patrick Lupardus Jianhuan Zhang Kelly M. Loyet Jawahar Sudhamsu Yan Wu
author_facet	Wei-Ching Liang Jianping Yin Patrick Lupardus Jianhuan Zhang Kelly M. Loyet Jawahar Sudhamsu Yan Wu
author_sort	Wei-Ching Liang
title	Dramatic activation of an antibody by a single amino acid change in framework
title_short	Dramatic activation of an antibody by a single amino acid change in framework
title_full	Dramatic activation of an antibody by a single amino acid change in framework
title_fullStr	Dramatic activation of an antibody by a single amino acid change in framework
title_full_unstemmed	Dramatic activation of an antibody by a single amino acid change in framework
title_sort	dramatic activation of an antibody by a single amino acid change in framework
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/0b851df1980f44c9ad9fd78eb3510ab2
work_keys_str_mv	AT weichingliang dramaticactivationofanantibodybyasingleaminoacidchangeinframework AT jianpingyin dramaticactivationofanantibodybyasingleaminoacidchangeinframework AT patricklupardus dramaticactivationofanantibodybyasingleaminoacidchangeinframework AT jianhuanzhang dramaticactivationofanantibodybyasingleaminoacidchangeinframework AT kellymloyet dramaticactivationofanantibodybyasingleaminoacidchangeinframework AT jawaharsudhamsu dramaticactivationofanantibodybyasingleaminoacidchangeinframework AT yanwu dramaticactivationofanantibodybyasingleaminoacidchangeinframework
_version_	1718418985110732800

Dramatic activation of an antibody by a single amino acid change in framework

Ejemplares similares