New Anti-Nodal Monoclonal Antibodies Targeting the Nodal Pre-Helix Loop Involved in Cripto-1 Binding

International Journal of Molecular Sciences

Annalia Focà³  Luca Sanguigno⁴  Giuseppina Focà³  Luigi Strizzi¹  Roberta Iannitti³  Rosanna Palumbo³  Mary J. C. Hendrix¹  Antonio Leonardi²  Menotti Ruvo³  Annamaria Sandomenico³ 

[1] Program in Cancer Biology and Epigenomics, Stanley Manne Children’s Research Institute at Ann and Robert H. Lurie Children’s Hospital of Chicago, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA; E-Mails:;Department of Molecular Medicine and Medical Biotechnologies, Università degli Studi di Napoli “Federico II”, Naples 80131, Italy;Institute of Biostructure and Bioimaging, National Research Council (IBB-CNR) and Centro Interuniversitario di Ricerca sui Peptidi Bioattivi (CIRPeB), Università degli Studi di Napoli “Federico II”, Naples 80134, Italy; E-Mails:;Bioker Multimedica, Naples 80131, Italy; E-Mail:

关键词: Fab fragments;    monoclonal antibody;    melanoma;    nodal;    SPR;   

DOI  :  10.3390/ijms160921342

来源: mdpi

【 摘 要 】

Nodal is a potent embryonic morphogen belonging to the TGF-β superfamily. Typically, it also binds to the ALK4/ActRIIB receptor complex in the presence of the co-receptor Cripto-1. Nodal expression is physiologically restricted to embryonic tissues and human embryonic stem cells, is absent in normal cells but re-emerges in several human cancers, including melanoma, breast, and colon cancer. Our aim was to obtain mAbs able to recognize Nodal on a major CBR (Cripto-Binding-Region) site and to block the Cripto-1-mediated signalling. To achieve this, antibodies were raised against hNodal(44–67) and mAbs generated by the hybridoma technology. We have selected one mAb, named 3D1, which strongly associates with full-length rhNodal (K_D 1.4 nM) and recognizes the endogenous protein in a panel of human melanoma cell lines by western blot and FACS analyses. 3D1 inhibits the Nodal-Cripto-1 binding and blocks Smad2/3 phosphorylation. Data suggest that inhibition of the Nodal-Cripto-1 axis is a valid therapeutic approach against melanoma and 3D1 is a promising and interesting agent for blocking Nodal-Cripto mediated tumor development. These findings increase the interest for Nodal as both a diagnostic and prognostic marker and as a potential new target for therapeutic intervention.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.