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Development of conformational antibodies targeting Cripto-1 with neutralizing effects in vitro

Giuseppina Focà1, Emanuela Iaccarino1, Annalia Focà1

  • 1Institute of Biostructure and Bioimaging, National Research Council (IBB-CNR), Naples, Italy.

Biochimie
|February 1, 2019
PubMed

Insights

Researchers developed novel monoclonal antibodies targeting the Cripto-1 protein

Area of Science:

  • Oncology
  • Developmental Biology
  • Biochemistry

Background:

  • Human Cripto-1 (Cripto-1), an EGF-CFC superfamily member, regulates embryonic development and oncogenesis.
  • Cripto-1 is absent in normal adult tissues but re-expressed in various tumors, making it a therapeutic target.
  • Blocking Cripto-1's CFC domain function is a promising strategy to inhibit tumor growth.

Purpose of the Study:

  • To generate and characterize murine monoclonal antibodies (mAbs) against the human Cripto-1 CFC domain.
  • To identify specific mAbs that recognize key residues essential for Cripto-1's interaction with ALK4 and GRP78.
  • To evaluate the potential of these mAbs as therapeutic and theranostic agents for cancer.

Main Methods:

  • Generation of murine monoclonal antibodies against a synthetic Cripto-1 CFC domain.
  • Subtractive ELISA, Surface Plasmon Resonance (SPR), and epitope mapping for antibody specificity.
  • Flow cytometry (FACS) to detect Cripto-1 on cancer cells.
  • In vitro cellular signaling and cell cycle analyses.

Main Results:

  • Successfully generated and characterized mAbs against the Cripto-1 CFC domain.
  • Identified specific mAbs recognizing conformational epitopes crucial for ALK4/GRP78 binding.
  • Demonstrated that the 1B4 antibody recognizes both membrane-bound and soluble Cripto-1 on human cancer cells.
  • Observed functional effects on cellular signaling and cell cycle, suggesting neutralization of oncogenic activity.

Conclusions:

  • Selected anti-CFC mAbs show potential to neutralize Cripto-1's oncogenic activity.
  • These mAbs can serve as theranostic agents for targeting Cripto-1-overexpressing cancers.
  • The developed antibodies may help overcome drug resistance in cancer therapies.
Keywords:
Cancer biomarkerCripto-1Neutralizing monoclonal antibodies

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