Cited by 72Monoclonal and Polyclonal Antibodies ResearchToxin Mechanisms and ImmunotoxinsHER2/EGFR in Cancer Research

Humanization of the Bispecific Epidermal Growth Factor Receptor × CD3 Diabody and Its Efficacy as a Potential Clinical Reagent

Abstract

Purpose: Bispecific antibodies (BsAb) have been exploited as both cancer immunodiagnostics and cancer therapeutics and show promise in clinical trials of cancer imaging and therapy. For development of BsAbs as clinical reagents, we have focused on construction of small recombinant BsAbs, called bispecific diabodies. Here, we constructed and characterized a humanized bispecific diabody. Experimental Design: We have reported significant antitumor activity of an anti-epidermal growth factor receptor (EGFR) × anti-CD3 bispecific diabody (Ex3) in in vitro cytotoxicity assays and in vivo. We humanized the Ex3 diabody (hEx3) by grafting the complementarity-determining region and compared its biological properties with those of Ex3. We also tested its physiologic stability and ability to alter survival in xenografted mice. Results: The final yield of hEx3 was 10 times that of Ex3, and refolded hEx3 and Ex3 showed identical binding profiles in EGFR-positive cell lines and EGFR-transfected Chinese hamster ovary cells. hEx3 showed dose-dependent cytotoxicity to EGFR-positive cell lines, which could be specifically inhibited by parental monoclonal antibody IgGs against EGFR or CD3 antigens. The heterodimeric structure was retained in PBS for 6 months, and growth inhibition was maintained after incubation under physiologic conditions. Coadministration of hEx3 with T-LAK cells and interleukin-2 prolonged the survival of nude mice with human colon carcinoma. Conclusions: The humanized diabody hEx3 is an attractive molecule for cancer therapy and may provide important insights into the development of EGFR-based cancer-targeting reagents.

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Crossref · 10.1158/1078-0432.ccr-06-0059

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Authors
Asano, R., Sone, Y., Makabe, K., Tsumoto, K., Hayashi, H., Katayose, Y., Unno, M., Kudo, T., & Kumagai, I.
Affiliations
  • 1Department of Biomolecular Engineering, Graduate School of Engineering;
  • 2Division of Gastroenterological Surgery, Department of Surgery, Graduate School of Medicine; and
  • Tohoku University (JP)
Venue
Clinical Cancer Research
Published
2006-07-01
Volume
12
Issue
13
Pages
4036-4042
ISSN
  • 1078-0432 (ISSN-L)
  • 1557-3265
OpenAlex
W2170996501
Cited by
72
References
46
Citations by year
2026: 3 · 2025: 4 · 2024: 4 · 2023: 3 · 2021: 2 · 2020: 4 · 2019: 2 · 2018: 4 · 2017: 1 · 2016: 1 · 2015: 1 · 2014: 3 · 2013: 3 · 2012: 7
FWCI
5.11
Type
journal article
Language
en
Open access
closed
Locations
  • Clinical Cancer Research (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
  • Life Sciences › Immunology and Microbiology › Immunology › Toxin Mechanisms and Immunotoxins
  • Health Sciences › Medicine › Oncology › HER2/EGFR in Cancer Research
Keywords
humanization, epidermal growth factor receptor, CD3, anticancer activity, cytotoxicity, protein stability, colon cancer, interleukin-2, cancer immunotherapy
MeSH terms
Animals, Antigen-Antibody Reactions, Antineoplastic Agents, Carcinoma, Cloning, Molecular, Colonic Neoplasms, Cricetulus, Disease Models, Animal, Dose-Response Relationship, Drug, Drug Screening Assays, Antitumor, Flow Cytometry, Cricetinae, Humans, Mice, Nude, Protein Binding, Transplantation, Heterologous, CHO Cells, CD3 Complex, Antibodies, Bispecific, Xenograft Model Antitumor Assays, Cell Line, Tumor, Cell Proliferation, Mice, ErbB Receptors

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