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A simple screening system for claudin binders using an scFv library derived from claudin‐immunized mice
Author(s) -
Kodaka Miki,
Takahashi Azusa,
Yamaura Toshiaki,
Kakamu Yohei,
Matsuhisa Koji,
Matsushita Kyohei,
Watari Akihiro,
Kondoh Masuo,
Yagi Kiyohito
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.620.12
Subject(s) - claudin , simple (philosophy) , tight junction , chemistry , biochemistry , philosophy , epistemology
Epithelial cells (ECs) separate the body from the external environment, and most malignant tumors are derived from epithelium. ECs are potent targets for drug absorption and cancer‐targeting. Tight junctions (TJs) are present between adjacent ECs. TJs prevent free movement of solutes across epithelial cell sheets and of transmembrane proteins between the apical and basolateral membranes. Claudin (CL), a family of tetra‐transmembrane proteins consisting of 24 members, is a key TJ‐sealing component and CLs are overexpressed in at least 12 human cancers. A CL4 binder modulated the TJ‐barrier, leading to enhancement of the mucosal absorption of biologics. These findings indicate that a CL binder may be useful for modulation of drug absorption and cancer‐targeting. However, few CL binders have been developed since CLs are weakly antigenic and their structures have not been determined. In this study, we aimed to develop a novel CL binder screening system using a CL‐displaying budded baculovirus (CL‐BV). CL3 KO mice were immunized with CL3‐BV, and an scFv library was prepared from the spleen. By screening the binding of the scFv library to CL‐BV, we identified several scFv clones, which bound to CL1, 3 or 4 with high affinity. These CL‐ligands reduced TJ‐integrity. These findings indicate that the scFv library derived from mice immunized with CL‐BV may be a promising screening system for identification of a ligand for CL.