Establishment of EMab-134, a Sensitive and Specific Anti-Epidermal Growth Factor Receptor Monoclonal Antibody for Detecting Squamous Cell Carcinoma Cells of the Oral Cavity

Shunsuke Itai, Shinji Yamada, Mika K. Kaneko, Yao Wen Chang, Hiroyuki Harada, Yukinari Kato

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Epidermal growth factor receptor (EGFR), a receptor tyrosine kinase, activates downstream signaling cascades in many tumors. In this study, we established novel anti-EGFR monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. We immunized mice with a combination of the extracellular domain of EGFR and EGFR-overexpressing LN229 glioblastoma cells (LN229/EGFR) and performed the first screening using enzyme-linked immunosorbent assay. Next, we selected mAbs using flow cytometry. Among 156 established clones, two mAbs, EMab-51 (IgG1, kappa) and EMab-134 (IgG1, kappa), reacted with EGFR in Western blot analysis; EMab-134 showed a much higher sensitivity compared with EMab-51. We compared the binding affinities of EMab-51 and EMab-134 using flow cytometry; the calculated KD values for EMab-51 and EMab-134 against SAS cells/HSC-2 cells were 9.2 × 10-9 M/9.9 × 10-9 M and 2.6 × 10-9 M/8.3 × 10-9 M, respectively, indicating that EMab-134 has a higher affinity to EGFR-expressing cells. Immunohistochemical analysis of EMab-51 and EMab-134 showed sensitive and specific reactions against oral cancer cells; EMab-134 demonstrated a much higher sensitivity (36/38 cases; 94.7%) to oral squamous cell carcinomas compared with EMab-51 (6/38 cases; 15.8%). This novel anti-EGFR mAb, EMab-134, could be advantageous for detecting EGFR in the pathological analysis of EGFR-expressing cancers.

Original languageEnglish
Pages (from-to)272-281
Number of pages10
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume36
Issue number6
DOIs
Publication statusPublished - 2017 Dec

Keywords

  • EGFR
  • immunohistochemistry
  • monoclonal antibody
  • oral cancer

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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