Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing

Abstract A major class of bispecific antibodies (BsAbs) utilizes heterodimeric Fc to produce the native immunoglobulin G (IgG) structure. Because appropriate pairing of heavy and light chains is required, the design of BsAbs produced through recombination or reassembly of two separately-expressed an...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Hiroki Akiba, Tomoko Ise, Satoshi Nagata, Haruhiko Kamada, Hiroaki Ohno, Kouhei Tsumoto
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8887dd3574c14623af5fb59364d6b6a2
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:8887dd3574c14623af5fb59364d6b6a2
record_format	dspace
spelling	oai:doaj.org-article:8887dd3574c14623af5fb59364d6b6a22021-12-02T17:17:40ZProduction of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing10.1038/s41598-021-98855-32045-2322https://doaj.org/article/8887dd3574c14623af5fb59364d6b6a22021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-98855-3https://doaj.org/toc/2045-2322Abstract A major class of bispecific antibodies (BsAbs) utilizes heterodimeric Fc to produce the native immunoglobulin G (IgG) structure. Because appropriate pairing of heavy and light chains is required, the design of BsAbs produced through recombination or reassembly of two separately-expressed antigen-binding fragments is advantageous. One such method uses intein-mediated protein trans-splicing (IMPTS) to produce an IgG1-based structure. An extra Cys residue is incorporated as a consensus sequence for IMPTS in successful examples, but this may lead to potential destabilization or disturbance of the assay system. In this study, we designed a BsAb linked by IMPTS, without the extra Cys residue. A BsAb binding to both TNFR2 and CD30 was successfully produced. Cleaved side product formation was inevitable, but it was minimized under the optimized conditions. The fine-tuned design is suitable for the production of IgG-like BsAb with high symmetry between the two antigen-binding fragments that is advantageous for screening BsAbs.Hiroki AkibaTomoko IseSatoshi NagataHaruhiko KamadaHiroaki OhnoKouhei TsumotoNature 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 Hiroki Akiba Tomoko Ise Satoshi Nagata Haruhiko Kamada Hiroaki Ohno Kouhei Tsumoto Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing
description	Abstract A major class of bispecific antibodies (BsAbs) utilizes heterodimeric Fc to produce the native immunoglobulin G (IgG) structure. Because appropriate pairing of heavy and light chains is required, the design of BsAbs produced through recombination or reassembly of two separately-expressed antigen-binding fragments is advantageous. One such method uses intein-mediated protein trans-splicing (IMPTS) to produce an IgG1-based structure. An extra Cys residue is incorporated as a consensus sequence for IMPTS in successful examples, but this may lead to potential destabilization or disturbance of the assay system. In this study, we designed a BsAb linked by IMPTS, without the extra Cys residue. A BsAb binding to both TNFR2 and CD30 was successfully produced. Cleaved side product formation was inevitable, but it was minimized under the optimized conditions. The fine-tuned design is suitable for the production of IgG-like BsAb with high symmetry between the two antigen-binding fragments that is advantageous for screening BsAbs.
format	article
author	Hiroki Akiba Tomoko Ise Satoshi Nagata Haruhiko Kamada Hiroaki Ohno Kouhei Tsumoto
author_facet	Hiroki Akiba Tomoko Ise Satoshi Nagata Haruhiko Kamada Hiroaki Ohno Kouhei Tsumoto
author_sort	Hiroki Akiba
title	Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing
title_short	Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing
title_full	Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing
title_fullStr	Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing
title_full_unstemmed	Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing
title_sort	production of igg1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8887dd3574c14623af5fb59364d6b6a2
work_keys_str_mv	AT hirokiakiba productionofigg1basedbispecificantibodywithoutextracysteineresidueviainteinmediatedproteintranssplicing AT tomokoise productionofigg1basedbispecificantibodywithoutextracysteineresidueviainteinmediatedproteintranssplicing AT satoshinagata productionofigg1basedbispecificantibodywithoutextracysteineresidueviainteinmediatedproteintranssplicing AT haruhikokamada productionofigg1basedbispecificantibodywithoutextracysteineresidueviainteinmediatedproteintranssplicing AT hiroakiohno productionofigg1basedbispecificantibodywithoutextracysteineresidueviainteinmediatedproteintranssplicing AT kouheitsumoto productionofigg1basedbispecificantibodywithoutextracysteineresidueviainteinmediatedproteintranssplicing
_version_	1718381155997188096

Production of IgG1-based bispecific antibody without extra cysteine residue via intein-mediated protein trans-splicing

Ejemplares similares