Isolation of scFvs to in vitro produced extracellular domains of EGFR family members

Cancer Biother Radiopharm. 2005 Dec;20(6):603-13. doi: 10.1089/cbr.2005.20.603.


The members of the epidermal growth factor receptor (EGFR) family are over expressed in a variety of malignancies and are frequently linked to aggressive disease and a poor prognosis. Although clinically effective monoclonal antibodies (MAbs) have been developed to target HER2 and EGFR, the remaining two family members, HER3 and HER4, have not been the subject of significant efforts. In this paper, we have taken the initial steps required to generate antibodies with potential clinically utility that target the members of the EGFR family. The genes for the extracellular domains (ECDs) of all four members of the EGFR family were cloned and used to stably transfect 293 (HEK) cells. Milligram quantities of each ECD were produced and characterized. The HER3, HER4, and EGFR ECDs were then employed as targets for the selection of antibodies from naïve human scFv (single-chain Fv) phage display libraries. Six unique scFv clones were isolated that bound specifically to HER3, 13 unique clones were isolated with specificity for HER4 and 52 unique anti-EGFR clones were isolated. These scFvs provide a valuable and potentially clinically relevant panel of agents to target the members of the EGFR family.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • Cell Line
  • Cell Line, Tumor
  • Cloning, Molecular
  • DNA Primers
  • ErbB Receptors / genetics*
  • Female
  • Humans
  • Immunoglobulin Variable Region / isolation & purification*
  • Kidney
  • Molecular Sequence Data
  • Ovarian Neoplasms
  • Receptor, ErbB-4
  • Transfection


  • DNA Primers
  • Immunoglobulin Variable Region
  • ERBB4 protein, human
  • ErbB Receptors
  • Receptor, ErbB-4