Mutation-specific antibodies for the detection of EGFR mutations in non-small-cell lung cancer

Clin Cancer Res. 2009 May 1;15(9):3023-8. doi: 10.1158/1078-0432.CCR-08-2739. Epub 2009 Apr 14.


Purpose: Activating mutations within the tyrosine kinase domain of epidermal growth factor receptor (EGFR) are found in approximately 10% to 20% of non-small-cell lung cancer (NSCLC) patients and are associated with response to EGFR inhibitors. The most common NSCLC-associated EGFR mutations are deletions in exon 19 and L858R mutation in exon 21, together accounting for 90% of EGFR mutations. To develop a simple, sensitive, and reliable clinical assay for the identification of EGFR mutations in NSCLC patients, we generated mutation-specific rabbit monoclonal antibodies against each of these two most common EGFR mutations and aimed to evaluate the detection of EGFR mutations in NSCLC patients by immunohistochemistry.

Experimental design: We tested mutation-specific antibodies by Western blot, immunofluorescence, and immunohistochemistry. In addition, we stained 40 EGFR genotyped NSCLC tumor samples by immunohistochemistry with these antibodies. Finally, with a panel of four antibodies, we screened a large set of NSCLC patient samples with unknown genotype and confirmed the immunohistochemistry results by DNA sequencing.

Results: These two antibodies specifically detect the corresponding mutant form of EGFR by Western blotting, immunofluorescence, and immunohistochemistry. Screening a panel of 340 paraffin-embedded NSCLC tumor samples with these antibodies showed that the sensitivity of the immunohistochemistry assay is 92%, with a specificity of 99% as compared with direct and mass spectrometry-based DNA sequencing.

Conclusions: This simple assay for detection of EGFR mutations in diagnostic human tissues provides a rapid, sensitive, specific, and cost-effective method to identify lung cancer patients responsive to EGFR-based therapies.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Biological Assay
  • Blotting, Western
  • Carcinoma, Non-Small-Cell Lung / genetics*
  • Carcinoma, Non-Small-Cell Lung / secondary
  • DNA Mutational Analysis
  • DNA, Neoplasm / genetics
  • DNA, Neoplasm / metabolism
  • ErbB Receptors / genetics*
  • Flow Cytometry
  • Humans
  • Immunoenzyme Techniques
  • Immunoglobulin G / immunology
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / pathology
  • Mice
  • Mice, Nude
  • Mutation / immunology*
  • Rabbits
  • Sensitivity and Specificity
  • Sequence Deletion
  • Transplantation, Heterologous
  • Tumor Cells, Cultured


  • Antibodies, Monoclonal
  • DNA, Neoplasm
  • Immunoglobulin G
  • EGFR protein, human
  • ErbB Receptors