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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
Summary
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.
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