Construction and characterization of single-chain antibodies against human insulin-like growth factor-I receptor from hybridomas producing 1H7 or 3B7 monoclonal antibody

Yu Kusada, Toru Morizono, Ayano Matsumoto-Takasaki, Keiko Sakai, Shuma Sato, Hideki Asanuma, Atsushi Takayanagi, Yoko Fujita-Yamaguchi

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Recombinant antibody consisting of the single-chain variable fragment (scFv) of 1H7 monoclonal antibody against insulin-like growth factor-I receptor (IGF-IR) and human IgG1 Fc domain, scFv-Fc, has been found to exhibit inhibitory effects on breast cancer growth in vitro and in vivo [Li et al. (2000) Cancer Immunol. Immunother. 49, 243; Sachdev et al. (2003) Cancer Res. 63, 627]. Various types of scFvs from hybridomas producing 1H7 or 3B7 mAb were constructed using conventional phage display technology to further characterize the specificity and affinity of anti-IGF-IR mAbs. Binding studies performed using either phage antibodies or soluble scFv proteins to IGF-IR or insulin receptor (IR) and IGF-IR pre-incubated with mAbs suggested that (i) 1H7 and 3B7 bind to IGF-IR but do not bind to its structurally related IR, (ii) either the VL-VH or VH-VL sequence order does not apparently affect specificity for IGF-IR and (iii) 1H7 and 3B7 bind the independent epitopes, located in or near the N-terminal (440-514) and C-terminal (62-184) domains of the α subunit, respectively. This study not only revealed new information on binding regions for two anti-IGF-IR mAbs, but also provided the scFv genes as tools for further manipulation of the affinity or development of new IGF-IR-targeted cancer therapeutics.

Original languageEnglish
Pages (from-to)9-19
Number of pages11
JournalJournal of biochemistry
Volume143
Issue number1
DOIs
Publication statusPublished - 2008 Jan 1

Keywords

  • Epitope specificity
  • Insulin receptor
  • Insulin-like growth factor I receptor
  • Phage display
  • Single-chain antibody

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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