A new approach to produce IgG4-like bispecific antibodies

Abstract While achieving rapid developments in recent years, bispecific antibodies are still difficult to design and manufacture, due to mispair of both heavy and light chains. Here we report a novel technology to make bispecific molecules. The knob-into-hole method was used to pair two distinct hea...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Caizhi Zhao, Wei Zhang, Guihua Gong, Liping Xie, Ming-Wei Wang, Youjia Hu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4c2d7b2d5b9d4fd3a54801a855b5b91f
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:4c2d7b2d5b9d4fd3a54801a855b5b91f
record_format	dspace
spelling	oai:doaj.org-article:4c2d7b2d5b9d4fd3a54801a855b5b91f2021-12-02T17:26:55ZA new approach to produce IgG4-like bispecific antibodies10.1038/s41598-021-97393-22045-2322https://doaj.org/article/4c2d7b2d5b9d4fd3a54801a855b5b91f2021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97393-2https://doaj.org/toc/2045-2322Abstract While achieving rapid developments in recent years, bispecific antibodies are still difficult to design and manufacture, due to mispair of both heavy and light chains. Here we report a novel technology to make bispecific molecules. The knob-into-hole method was used to pair two distinct heavy chains as a heterodimer. IgG4 S228P CH1-CL interface was then partially replaced by T-cell receptor α/β constant domain to increase the efficiency of cognate heavy and light chain pairing. Following expression and purification, the bispecific antibody interface exchange was confirmed by Western blotting and LC–MS/MS. To ensure its validity, we combined a monovalent bispecific antibody against PD-1 (sequence from Pembrolizumab) and LAG3 (sequence from Relatlimab). The results showed that the molecule could be assembled correctly at a ratio of 95% in cells. In vitro functional assay demonstrated that the purified bispecific antibody exhibits an enhanced agonist activity compared to that of the parental antibodies. Low immunogenicity was predicted by an open-access software and ADA test.Caizhi ZhaoWei ZhangGuihua GongLiping XieMing-Wei WangYoujia HuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Caizhi Zhao Wei Zhang Guihua Gong Liping Xie Ming-Wei Wang Youjia Hu A new approach to produce IgG4-like bispecific antibodies
description	Abstract While achieving rapid developments in recent years, bispecific antibodies are still difficult to design and manufacture, due to mispair of both heavy and light chains. Here we report a novel technology to make bispecific molecules. The knob-into-hole method was used to pair two distinct heavy chains as a heterodimer. IgG4 S228P CH1-CL interface was then partially replaced by T-cell receptor α/β constant domain to increase the efficiency of cognate heavy and light chain pairing. Following expression and purification, the bispecific antibody interface exchange was confirmed by Western blotting and LC–MS/MS. To ensure its validity, we combined a monovalent bispecific antibody against PD-1 (sequence from Pembrolizumab) and LAG3 (sequence from Relatlimab). The results showed that the molecule could be assembled correctly at a ratio of 95% in cells. In vitro functional assay demonstrated that the purified bispecific antibody exhibits an enhanced agonist activity compared to that of the parental antibodies. Low immunogenicity was predicted by an open-access software and ADA test.
format	article
author	Caizhi Zhao Wei Zhang Guihua Gong Liping Xie Ming-Wei Wang Youjia Hu
author_facet	Caizhi Zhao Wei Zhang Guihua Gong Liping Xie Ming-Wei Wang Youjia Hu
author_sort	Caizhi Zhao
title	A new approach to produce IgG4-like bispecific antibodies
title_short	A new approach to produce IgG4-like bispecific antibodies
title_full	A new approach to produce IgG4-like bispecific antibodies
title_fullStr	A new approach to produce IgG4-like bispecific antibodies
title_full_unstemmed	A new approach to produce IgG4-like bispecific antibodies
title_sort	new approach to produce igg4-like bispecific antibodies
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4c2d7b2d5b9d4fd3a54801a855b5b91f
work_keys_str_mv	AT caizhizhao anewapproachtoproduceigg4likebispecificantibodies AT weizhang anewapproachtoproduceigg4likebispecificantibodies AT guihuagong anewapproachtoproduceigg4likebispecificantibodies AT lipingxie anewapproachtoproduceigg4likebispecificantibodies AT mingweiwang anewapproachtoproduceigg4likebispecificantibodies AT youjiahu anewapproachtoproduceigg4likebispecificantibodies AT caizhizhao newapproachtoproduceigg4likebispecificantibodies AT weizhang newapproachtoproduceigg4likebispecificantibodies AT guihuagong newapproachtoproduceigg4likebispecificantibodies AT lipingxie newapproachtoproduceigg4likebispecificantibodies AT mingweiwang newapproachtoproduceigg4likebispecificantibodies AT youjiahu newapproachtoproduceigg4likebispecificantibodies
_version_	1718380785841471488

A new approach to produce IgG4-like bispecific antibodies

Ejemplares similares