title: Developing and characterization of single chain variable fragment (scFv) antibody against frizzled 7 (Fzd7) receptor.
abstract: ABSTACT Wnt/β-catenin signaling pathway through Frizzled receptors has been shown to play a key role in both normal development and tumorigenesis. Overexpression of Wnt pathway genes, such as Fzd7 in several malignancies is well-documented. Therefore, targeting of Fzd7 and its ligand inhibits cancer cells proliferation metastasis. In the present study we isolated single chain variable fragments (scFvs) against Fzd7 receptor using phage display method. Semi-synthetic human naive antibody libraries (Tomlinson I + J) was employed in panning procedure to isolate specific scFv against specific peptide from extracellular domain of Fzd7 receptor. The reactivity and growth inhibition effects of the selected antibodies was evaluated using enzyme-linked immunosorbent assay (ELISA), MTT and annexin V assays, respectively. Seven scFvs reactive to Fzd7 were selected following 4 rounds of panning. The results showed that the selected scFvs inhibits cell growth through apoptosis cell death in a triple negative breast cancer cells, MDA-MB-231. Given that Fzd7 and Wnt pathway plays a critical role in tumor progression, selected blocking scFvs represent significant potential for immunotherapy of breast cancer cells.
authors.affiliation: a Immunology Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|b Drug Applied Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|c Department of Immunology , School of Medicine, Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Hamid
authors.initials: H
authors.lastname: Nickho
authors.affiliation: d Pishtaz Teb Zaman Diagnostics , Tehran , Iran.|e Department of Laboratory Sciences , Faculty of Paramedical Sciences, Alborz University of Medical Sciences , Karaj , Iran.
authors.forename: Vahid
authors.initials: V
authors.lastname: Younesi
authors.affiliation: a Immunology Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|b Drug Applied Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|c Department of Immunology , School of Medicine, Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Leili
authors.initials: L
authors.lastname: Aghebati-Maleki
authors.affiliation: a Immunology Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|b Drug Applied Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|c Department of Immunology , School of Medicine, Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Morteza
authors.initials: M
authors.lastname: Motallebnezhad
authors.affiliation: a Immunology Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|b Drug Applied Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|c Department of Immunology , School of Medicine, Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Jafar
authors.initials: J
authors.lastname: Majidi Zolbanin
authors.affiliation: a Immunology Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|b Drug Applied Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|c Department of Immunology , School of Medicine, Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Aliakbar
authors.initials: A
authors.lastname: Movassagh Pour
authors.affiliation: b Drug Applied Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.|c Department of Immunology , School of Medicine, Tabriz University of Medical Sciences , Tabriz , Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Yousefi
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D045744:Cell Line, Tumor; D061505:Cell Surface Display Techniques; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D051157:Frizzled Receptors / Q000008:administration & dosage* / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D057127:Single-Chain Antibodies / Q000008:administration & dosage* / Q000276:immunology*; D016896:Treatment Outcome; D064726:Triple Negative Breast Neoplasms / Q000188:drug therapy* / Q000276:immunology* / Q000473:pathology
chemical_list: C115810:FZD7 protein, human; D051157:Frizzled Receptors; D057127:Single-Chain Antibodies
keywords: Fzd7; Wnt; breast cancer; phage display; scFv