1. Isolation and preliminary characterization of a human 'phage display'-derived antibody against neural adhesion molecule-1 antigen interfering with fibroblast growth factor receptor-1 binding
- Author
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Maria Luisa Dupuis, Alessandra Boe, Mauro Andreotti, Carla Raggi, Mara Gellini, Alessandra Mallano, Alessandro Ascione, Eleonora Petrucci, Gianni Colotti, Stefano Barca, Stefano Vella, Roberta Riccioni, and Michela Flego
- Subjects
0301 basic medicine ,epithelial ovarian cancer ,Phage display ,Immunology ,Immunoglobulins ,Fibroblast growth factor ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,Immunology and Allergy ,Single-chain variable fragment ,Humans ,single-chain variable fragment ,Bacteriophages ,NCAM ,Neural Cell Adhesion Molecules ,Linear epitope ,biology ,Chemistry ,Fibroblast growth factor receptor 1 ,General Medicine ,Cell biology ,030104 developmental biology ,FGFR1 ,nervous system ,030220 oncology & carcinogenesis ,biology.protein ,Neural cell adhesion molecule ,Antibody ,phage display ,Protein Binding ,Signal Transduction - Abstract
BACKGROUND: The NCAM or CD56 antigen is a cell surface glycoprotein belonging to the immunoglobulin super-family involved in cell-cell and cell-matrix adhesion. NCAM is also over-expressed in many tumour types and is considered a tumour associated antigen, even if its role and biological mechanisms implicated in tumour progression and metastasis have not yet to be elucidated. In particular, it is quite well documented the role of the interaction between the NCAM protein and the fibroblast growth factor receptor-1 in metastasis and invasion, especially in the ovarian cancer progression. OBJECTIVE: Here we describe the isolation and preliminary characterization of a novel human anti-NCAM single chain Fragment variable antibody able to specifically bind NCAM-expressing cells, including epithelial ovarian cancer cells. METHODS: The antibody was isolate by phage display selection and was characterized by ELISA, FACS analysis and SPR experiments. Interference in EOC migration was analyzed by scratch test. RESULTS: It binds a partially linear epitope lying in the membrane proximal region of two fibronectin-like domains with a dissociation constant of 3.43 × 10-8 M. Interestingly, it was shown to interfere with the NCAM-FGFR1 binding and to partially decrease migration of EOC cells. CONCLUSIONS: According to our knowledge, this is the first completely human antibody able to interfere with this newly individuated cancer mechanism.
- Published
- 2020