Generation and characterization of a bispecific diabody targeting both EPH receptor A10 and CD3.

The EPH receptor A10 (EphA10) is up-regulated in breast cancer but is not normally expressed in healthy tissue, thus it has been suggested that EphA10 may be a useful target for cancer therapy. This study reports a diabody, an antibody derivative binding two different target molecules, EphA10 expressed in tumor cells and CD3 expressed in T cells, which showed T cell dependent-cytotoxicity. The diabody, which has His-tagged and FLAG-tagged chains, was expressed in Escherichia coli and purified in both heterodimer (Db-1) and homodimer (Db-2) formulations by liquid chromatography. Flow cytometry analysis using EphA10-expressing cells showed that binding activity of heterodimers was stronger than that of homodimers. Addition of diabodies to PBMC cultures resulted in T-cell mediated redirected lysis, and the bioactivity was consistent with the stronger binding activity of heterodimeric diabody formulations. Our results indicate that diabodies recognizing both EphA10 and CD3 could have a range of potential applications in cancer therapy, such as breast cancers that express the EPH receptor A10, especially triple negative breast cancer. Copyright © 2014 Elsevier Inc. All rights reserved.

Citation

Haruhiko Kamada, Shintaro Taki, Kazuya Nagano, Masaki Inoue, Daisuke Ando, Yohei Mukai, Kazuma Higashisaka, Yasuo Yoshioka, Yasuo Tsutsumi, Shin-ichi Tsunoda. Generation and characterization of a bispecific diabody targeting both EPH receptor A10 and CD3. Biochemical and biophysical research communications. 2015 Jan 24;456(4):908-12

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PMID: 25528586

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