Strategies to structure specific epitopes and antibodies for a membrane type protease that is involved in the metastasis.

초록

Epithin, a type II transmembrane serine protease is upregulated in a variety of epithelial cancer cells and tumor tissues. Its high level expression is implicated in the poor clinical outcome. It cleaves ECM proteins and activates many well known tumor associated proteins. We showed earlier that it is involved in angiogenesis, TGFβ induced EMT, transedothelial migration, and metastasis of epithelial cancers. We selected the structure specific epitopes of high antigenicity by structure modeling. The selected epitopes were tested in prevention of metastasis using the two breast cancer cell lines and their immunocompetent syngenic mouse hosts, Balb/c and C57BL6. Injecting cancer cells into tail veins after Immunization of conjugated peptides in adjuvants reduced metastasis significantly in the sequence specific fashion. Antisera obtained from the mice showed inhibitory activities in various cell based assays. We now established multiple strategies to screen the structure specific antibodies in order to avoid nonspecific cross reactive antibodies using multiple advanced/modified technologies. Polyclonal antisera that are raised using designed antigens were successfully tested for the presence of structure specific antibodies