Molecular Medicine

, Volume 15, Issue 1–2, pp 11–20 | Cite as

Rational Optimization of a Bispecific Ligand Trap Targeting EGF Receptor Family Ligands

  • Pei Jin
  • Juan Zhang
  • Malgorzata Beryt
  • Lisa Turin
  • Cathleen Brdlik
  • Ying Feng
  • Xiaomei Bai
  • Jim Liu
  • Brett Jorgensen
  • H. Michael Shepard
Research Article


The human epidermal growth factor (EGF) receptor (HER) family members cooperate in malignancy. Of this family, HER2 does not bind growth factors and HER3 does not encode an active tyrosine kinase. This diversity creates difficulty in creating pan-specific therapeutic HER family inhibitors. We have identified single amino acid changes in epidermal growth factor receptor (EGFR) and HER3 which create high affinity sequestration of the cognate ligands, and may be used as receptor decoys to downregulate aberrant HER family activity. In silico modeling and high throughput mutagenesis were utilized to identify receptor mutants with very high ligand binding activity. A single mutation (T15S; EGFR subdomain I) enhanced affinity for EGF (two-fold), TGF-α (twenty-six-fold), and heparin-binding (HB)-EGF (six-fold). This indicates that T15 is an important, previously undescribed, negative regulatory amino acid for EGFR ligand binding. Another mutation (Y246A; HER 3 subdomain II) enhanced neuregulin (NRG)1-β binding eight-fold, probably by interfering with subdomain II–IV interactions. Further work revealed that the HER3 subunit of an EGFR:HER3 heterodimer suppresses EGFR ligand binding. Optimization required reversing this suppression by mutation of the EGFR tether domain (G564A; subdomain IV). This mutation resulted in enhanced ligand binding (EGF, ten-fold; TGF-α, thirty-four-fold; HB-EGF, seventeen-fold; NRG1-β, thirty-one-fold). This increased ligand binding was reflected in improved inhibition of in vitro tumor cell proliferation and tumor suppression in a human non-small cell lung cancer xenograft model. In conclusion, amino acid substitutions were identified in the EGFR and HER3 ECDs that enhance ligand affinity, potentially enabling a pan-specific therapeutic approach for downregulating the HER family in cancer.



We thank Drs. Jay Sarup, Douglas Kawahara, and Dan Maneval for helpful discussions on this research, and Scott Patton for excellent editorial assistance.

Supplementary material

10020_2009_15010011_MOESM1_ESM.pdf (276 kb)
Rational Optimization of a Bispecific Ligand Trap Targeting EGF Receptor Family Ligands


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Copyright information

© The Feinstein Institute for Medical Research 2009

Authors and Affiliations

  • Pei Jin
    • 1
  • Juan Zhang
    • 1
  • Malgorzata Beryt
    • 1
  • Lisa Turin
    • 1
  • Cathleen Brdlik
    • 1
  • Ying Feng
    • 1
  • Xiaomei Bai
    • 1
  • Jim Liu
    • 1
  • Brett Jorgensen
    • 1
  • H. Michael Shepard
    • 1
  1. 1.Receptor BioLogix, Inc.Palo AltoUSA

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