Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528

Ryutaro Asano; Yukiko Sone; Keiko Ikoma; Hiroki Hayashi; Takeshi Nakanishi; Mitsuo Umetsu; Yu Katayose; Michiaki Unno; Toshio Kudo; Izumi Kumagai

doi:10.1093/protein/gzn037

Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528

Ryutaro Asano, Yukiko Sone, Keiko Ikoma, Hiroki Hayashi, Takeshi Nakanishi, Mitsuo Umetsu, Yu Katayose, Michiaki Unno, Toshio Kudo, Izumi Kumagai

Tohoku University

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

13 Citations (Scopus)

Abstract

We report the utility of in vitro refolding in the preparation of monomorphous hEx3 bispecific diabodies with epidermal growth factor receptor and CD3 retargeting from insoluble aggregates in Escherichia coli. Appropriate interaction between cognate variable heavy and light chains led to the formation of functional hEx3 heterodimers in a diabody format rather than inactive homodimers. The refolded hEx3 was found to exhibit almost the equivalent activity to the hEx3 and single-chain hEx3 (hEx3-scDb) prepared in a mammalian secretion system. We suggest that the preparation of hEx3 from bacterial insoluble material by means of in vitro refolding would be useful for industrial-scale production of the diabody for its potential use in clinical studies.

Original language	English
Pages (from-to)	597-603
Number of pages	7
Journal	Protein Engineering, Design and Selection
Volume	21
Issue number	10
DOIs	https://doi.org/10.1093/protein/gzn037
Publication status	Published - 2008 Oct

Keywords

Bispecific diabody
Cancer immunotherapy
EGFR
In vitro refolding
Small recombinant antibody

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1093/protein/gzn037

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title = "Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528",

abstract = "We report the utility of in vitro refolding in the preparation of monomorphous hEx3 bispecific diabodies with epidermal growth factor receptor and CD3 retargeting from insoluble aggregates in Escherichia coli. Appropriate interaction between cognate variable heavy and light chains led to the formation of functional hEx3 heterodimers in a diabody format rather than inactive homodimers. The refolded hEx3 was found to exhibit almost the equivalent activity to the hEx3 and single-chain hEx3 (hEx3-scDb) prepared in a mammalian secretion system. We suggest that the preparation of hEx3 from bacterial insoluble material by means of in vitro refolding would be useful for industrial-scale production of the diabody for its potential use in clinical studies.",

keywords = "Bispecific diabody, Cancer immunotherapy, EGFR, In vitro refolding, Small recombinant antibody",

author = "Ryutaro Asano and Yukiko Sone and Keiko Ikoma and Hiroki Hayashi and Takeshi Nakanishi and Mitsuo Umetsu and Yu Katayose and Michiaki Unno and Toshio Kudo and Izumi Kumagai",

note = "Funding Information: Health Sciences of the National Institute of Biomedical Innovation (07-21); Japan Society for the Promotion of Science (16106011 to I.K., 19760548 to R.A.).",

year = "2008",

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doi = "10.1093/protein/gzn037",

language = "English",

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T1 - Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528

AU - Asano, Ryutaro

AU - Sone, Yukiko

AU - Ikoma, Keiko

AU - Hayashi, Hiroki

AU - Nakanishi, Takeshi

AU - Umetsu, Mitsuo

AU - Katayose, Yu

AU - Unno, Michiaki

AU - Kudo, Toshio

AU - Kumagai, Izumi

N1 - Funding Information: Health Sciences of the National Institute of Biomedical Innovation (07-21); Japan Society for the Promotion of Science (16106011 to I.K., 19760548 to R.A.).

PY - 2008/10

Y1 - 2008/10

N2 - We report the utility of in vitro refolding in the preparation of monomorphous hEx3 bispecific diabodies with epidermal growth factor receptor and CD3 retargeting from insoluble aggregates in Escherichia coli. Appropriate interaction between cognate variable heavy and light chains led to the formation of functional hEx3 heterodimers in a diabody format rather than inactive homodimers. The refolded hEx3 was found to exhibit almost the equivalent activity to the hEx3 and single-chain hEx3 (hEx3-scDb) prepared in a mammalian secretion system. We suggest that the preparation of hEx3 from bacterial insoluble material by means of in vitro refolding would be useful for industrial-scale production of the diabody for its potential use in clinical studies.

AB - We report the utility of in vitro refolding in the preparation of monomorphous hEx3 bispecific diabodies with epidermal growth factor receptor and CD3 retargeting from insoluble aggregates in Escherichia coli. Appropriate interaction between cognate variable heavy and light chains led to the formation of functional hEx3 heterodimers in a diabody format rather than inactive homodimers. The refolded hEx3 was found to exhibit almost the equivalent activity to the hEx3 and single-chain hEx3 (hEx3-scDb) prepared in a mammalian secretion system. We suggest that the preparation of hEx3 from bacterial insoluble material by means of in vitro refolding would be useful for industrial-scale production of the diabody for its potential use in clinical studies.

KW - Bispecific diabody

KW - Cancer immunotherapy

KW - EGFR

KW - In vitro refolding

KW - Small recombinant antibody

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DO - 10.1093/protein/gzn037

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IS - 10

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Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528

Abstract

Keywords

UN SDGs

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