title: New Anti-Nodal Monoclonal Antibodies Targeting the Nodal Pre-Helix Loop Involved in Cripto-1 Binding.
abstract: 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 (KD 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.
authors.affiliation: 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. annalia.foca@gmail.com.|Department of Pharmacy, Università degli Studi di Napoli "Federico II", Naples 80131, Italy. annalia.foca@gmail.com.
authors.forename: Annalia
authors.initials: A
authors.lastname: Focà
authors.affiliation: Bioker Multimedica, Naples 80131, Italy. Luca.Sanguigno@multimedica.it.
authors.forename: Luca
authors.initials: L
authors.lastname: Sanguigno
authors.affiliation: 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. giuseppina.foca@gmail.com.|Department of Pharmacy, Università degli Studi di Napoli "Federico II", Naples 80131, Italy. giuseppina.foca@gmail.com.
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Focà
authors.affiliation: 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. LStrizzi@luriechildrens.org.
authors.forename: Luigi
authors.initials: L
authors.lastname: Strizzi
authors.affiliation: 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. robertaiannitti@gmail.com.
authors.forename: Roberta
authors.initials: R
authors.lastname: Iannitti
authors.affiliation: 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. rosanna.palumbo@cnr.it.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Palumbo
authors.affiliation: 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. m-hendrix@northwestern.edu.
authors.forename: Mary J C
authors.initials: MJ
authors.lastname: Hendrix
authors.affiliation: Department of Molecular Medicine and Medical Biotechnologies, Università degli Studi di Napoli "Federico II", Naples 80131, Italy. leonardi@unina.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: Leonardi
authors.affiliation: 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. menotti.ruvo@unina.it.
authors.forename: Menotti
authors.initials: M
authors.lastname: Ruvo
authors.affiliation: 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. annamaria.sandomenico@gmail.com.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Sandomenico
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000494:pharmacology; D018604:Epitope Mapping / Q000379:methods; D000939:Epitopes / Q000737:chemistry / Q000378:metabolism; D058851:GPI-Linked Proteins / Q000737:chemistry / Q000378:metabolism; D055412:Growth Differentiation Factors / Q000737:chemistry; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism; D055457:Nodal Protein / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D058851:GPI-Linked Proteins; D055412:Growth Differentiation Factors; D007140:Immunoglobulin Fab Fragments; D036341:Intercellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; D055457:Nodal Protein; D010455:Peptides; C070636:TDGF1 protein, human
keywords: Fab fragments; SPR; melanoma; monoclonal antibody; nodal