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

干扰成纤维细胞生长因子受体-1 结合的抗神经粘附分子-1 抗原的人类“噬菌体展示”衍生抗体的分离和初步表征

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作者:Michela Flego, Gianni Colotti, Alessandro Ascione, Maria Luisa Dupuis, Eleonora Petrucci, Roberta Riccioni, Mauro Andreotti, Carla Raggi, Alessandra Boe, Stefano Barca, Mara Gellini, Stefano Vella, Alessandra Mallano

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.

Conclusions

According to our knowledge, this is the first completely human antibody able to interfere with this newly individuated cancer mechanism.

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.

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.

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.

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