Generation of camelid VHH bispecific constructs via in-cell intein-mediated protein trans-splicing

Yuki Shibuya; Natsuki Haga; Ryutaro Asano; Hikaru Nakazawa; Takamitsu Hattori; Daisuke Takeda; Aruto Sugiyama; Reiko Kurotani; Izumi Kumagai; Mitsuo Umetsu; Koki Makabe

Generation of camelid VHH bispecific constructs via in-cell intein-mediated protein trans-splicing

Yuki Shibuya, Natsuki Haga, Ryutaro Asano, Hikaru Nakazawa, Takamitsu Hattori, Daisuke Takeda, Aruto Sugiyama, Reiko Kurotani, Izumi Kumagai, Mitsuo Umetsu, Koki Makabe

ENG - Biomolecular Engineering

Research output: Contribution to journal › Article › peer-review

6 Citations (Scopus)

Abstract

Production of various combinations of bispecific variable domain of heavy chain of heavy chain-only antibody (VHH) constructs to evaluate their therapeutic potential usually requires several gene-engineering steps. Here, we present an alternative method of creating bispecific VHH constructs in vivo through protein trans-splicing (PTS) reaction; this method may reduce the number of gene manipulation steps required. As a proof-of-concept, we constructed a bispecific antibody (bsAb) containing an anti-epidermal growth factor receptor VHH and anti-green fluorescent protein VHH, and we evaluated and confirmed its bispecificity. We also tested antibody labeling by fluorescent protein tagging using the PTS reaction. Compared with the conventional gene construction method, bsAb construction via PTS is a promising alternative approach for generating multiple bsAb combinations.

Original language	English
Pages (from-to)	15-21
Number of pages	7
Journal	Protein Engineering, Design and Selection
Volume	30
Issue number	1
Publication status	Published - 2017

Keywords

Bispecific antibody
Intein
Protein splicing

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biochemistry
Molecular Biology

Cite this

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title = "Generation of camelid VHH bispecific constructs via in-cell intein-mediated protein trans-splicing",

abstract = "Production of various combinations of bispecific variable domain of heavy chain of heavy chain-only antibody (VHH) constructs to evaluate their therapeutic potential usually requires several gene-engineering steps. Here, we present an alternative method of creating bispecific VHH constructs in vivo through protein trans-splicing (PTS) reaction; this method may reduce the number of gene manipulation steps required. As a proof-of-concept, we constructed a bispecific antibody (bsAb) containing an anti-epidermal growth factor receptor VHH and anti-green fluorescent protein VHH, and we evaluated and confirmed its bispecificity. We also tested antibody labeling by fluorescent protein tagging using the PTS reaction. Compared with the conventional gene construction method, bsAb construction via PTS is a promising alternative approach for generating multiple bsAb combinations.",

keywords = "Bispecific antibody, Intein, Protein splicing",

author = "Yuki Shibuya and Natsuki Haga and Ryutaro Asano and Hikaru Nakazawa and Takamitsu Hattori and Daisuke Takeda and Aruto Sugiyama and Reiko Kurotani and Izumi Kumagai and Mitsuo Umetsu and Koki Makabe",

note = "Funding Information: This study was supported by JSPS KAKENHI Grants number 26670051 and 24000011, and Kurata Grant from the Kurata Memorial Hitachi Science and Technology Foundation. Publisher Copyright: {\textcopyright} The Author 2016. Published by Oxford University Press. All rights reserved.",

year = "2017",

language = "English",

volume = "30",

pages = "15--21",

journal = "Protein Engineering, Design and Selection",

issn = "1741-0126",

publisher = "Oxford University Press",

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TY - JOUR

T1 - Generation of camelid VHH bispecific constructs via in-cell intein-mediated protein trans-splicing

AU - Shibuya, Yuki

AU - Haga, Natsuki

AU - Asano, Ryutaro

AU - Nakazawa, Hikaru

AU - Hattori, Takamitsu

AU - Takeda, Daisuke

AU - Sugiyama, Aruto

AU - Kurotani, Reiko

AU - Kumagai, Izumi

AU - Umetsu, Mitsuo

AU - Makabe, Koki

N1 - Funding Information: This study was supported by JSPS KAKENHI Grants number 26670051 and 24000011, and Kurata Grant from the Kurata Memorial Hitachi Science and Technology Foundation. Publisher Copyright: © The Author 2016. Published by Oxford University Press. All rights reserved.

PY - 2017

Y1 - 2017

N2 - Production of various combinations of bispecific variable domain of heavy chain of heavy chain-only antibody (VHH) constructs to evaluate their therapeutic potential usually requires several gene-engineering steps. Here, we present an alternative method of creating bispecific VHH constructs in vivo through protein trans-splicing (PTS) reaction; this method may reduce the number of gene manipulation steps required. As a proof-of-concept, we constructed a bispecific antibody (bsAb) containing an anti-epidermal growth factor receptor VHH and anti-green fluorescent protein VHH, and we evaluated and confirmed its bispecificity. We also tested antibody labeling by fluorescent protein tagging using the PTS reaction. Compared with the conventional gene construction method, bsAb construction via PTS is a promising alternative approach for generating multiple bsAb combinations.

AB - Production of various combinations of bispecific variable domain of heavy chain of heavy chain-only antibody (VHH) constructs to evaluate their therapeutic potential usually requires several gene-engineering steps. Here, we present an alternative method of creating bispecific VHH constructs in vivo through protein trans-splicing (PTS) reaction; this method may reduce the number of gene manipulation steps required. As a proof-of-concept, we constructed a bispecific antibody (bsAb) containing an anti-epidermal growth factor receptor VHH and anti-green fluorescent protein VHH, and we evaluated and confirmed its bispecificity. We also tested antibody labeling by fluorescent protein tagging using the PTS reaction. Compared with the conventional gene construction method, bsAb construction via PTS is a promising alternative approach for generating multiple bsAb combinations.

KW - Bispecific antibody

KW - Intein

KW - Protein splicing

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M3 - Article

C2 - 27881685

AN - SCOPUS:85021858670

SN - 1741-0126

VL - 30

SP - 15

EP - 21

JO - Protein Engineering, Design and Selection

JF - Protein Engineering, Design and Selection

IS - 1

ER -

Generation of camelid VHH bispecific constructs via in-cell intein-mediated protein trans-splicing

Abstract

Keywords

ASJC Scopus subject areas

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