Development of conformational antibodies targeting Cripto-1 with neutralizing effects in vitro(199 visite) Foca G, Iaccarino E, Foca A, Sanguigno L, Untiveros G, Cuevas-nunez M, Strizzi L, Leonardi A, Ruvo M, Sandomenico A
Impact factor: 3.787, Impact factor a 5 anni: 3.728
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Parole chiave: Activin Receptors, Type I, Immunology, Metabolism, Animals, Antibodies, Monoclonal, Murine-Derived, Chemistry, Neoplasm, Neutralizing, Cell Line, Tumor, Flow Cytometry, Gpi-Linked Proteins, Heat-Shock Proteins, Humans, Intercellular Signaling Peptides And Proteins, Inbred Balb C, Neoplasm Proteins, Protein Domains, Cancer Biomarker, Cripto-1, Neutralizing Monoclonal Antibodies, Type I Immunology Metabolism
, Murine-Derived Chemistry Immunology
, Neoplasm Chemistry Immunology
, Neutralizing Chemistry Immunology
, Gpi-Linked Proteins Immunology Metabolism
, Heat-Shock Proteins Immunology Metabolism
, Intercellular Signaling Peptides And Proteins Immunology Metabolism
, Mice
, Neoplasm Proteins Immunology Metabolism,
Affiliazioni:
*** IBB - CNR *** Institute of Biostructure and Bioimaging, National Research Council (IBB-CNR), Naples, Italy., Department of Molecular Medicine and Medical Biotechnology, University of Naples "Federico II", Naples, Italy., Midwestern University, Colleges of Graduate Studies, Dwners Grove, Chicago, IL, USA., Midwestern University, Colleges of Graduate Studies, Dwners Grove, Chicago, IL, USA; College of Dental Medicine, Dwners Grove, Chicago, IL, USA., Institute of Biostructure and Bioimaging, National Research Council (IBB-CNR), Naples, Italy. Electronic address: menotti.ruvo@unina.it., Institute of Biostructure and Bioimaging, National Research Council (IBB-CNR), Naples, Italy. Electronic address: annamaria.sandomenico@cnr.it.,
Riferimenti:
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Human Cripto-1 (Cripto-1), the founding member of the EGF-CFC superfamily, is a key regulator of many processes during embryonic development and oncogenesis. Cripto-1 is barely present or even absent in normal adult tissues while it is aberrantly re-expressed in various tumors. Blockade of the CFC domain-mediated Cripto-1 functions is acknowledged as a promising therapeutic intervention point to inhibit the tumorigenic activity of the protein. In this work, we report the generation and characterization of murine monoclonal antibodies raised against the synthetic folded CFC [112-150] domain of the human protein. Through subtractive ELISA assays clones were screened for the ability to specifically recognize "hot spot" residues on the CFC domain, which are crucial for the interaction with Activin Type I receptor (ALK4) and GRP78. On selected antibodies, SPR and epitope mapping studies have confirmed their specificity and have revealed that recognition occurs only on a conformational epitope. Furthermore, FACS analyses have confirmed the ability of 1B4 antibody to recognize the membrane-anchored and soluble native Cripto-1 protein in a panel of human cancer cells. Finally, we have evaluated its functional effects through in vitro cellular signaling assays and cell cycle analysis. These findings suggest that the selected anti-CFC mAbs have the potential to neutralize the protein oncogenic activity and may be used as theranostic molecules suitable as tumor homing agents for Cripto-1-overexpressing cancer cells and tissues and to overcome drug-resistance in routine cancer therapies.
Ciccarelli M, Sorriento D, Coscioni E, Iaccarino G, Santulli G * Adrenergic Receptors(30 visite) Endocrinol Of The Heart In Health And Dis (ISSN: 9780-1280311249780128031117), 2016; N/D: 285-315. Impact Factor:0 DettagliEsporta in BibTeXEsporta in EndNote
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