Attempt of Cancer Pain Control with the aid of Anti-TRPV Channel Antibody

点击翻译按钮获取中文摘要

This project intends to produce a new therapeutic antibody that would ameliorate protracted cancer pain syndrome by utilizing a new finding that a cation channel called TRPV1 constitutes one of the final pathway of signaling pain sensation in neuronal cells. Although the polyclonal antibody raised against the bacterially produced TRPV1 E3 bop recognized the antigen protein with a high titer in Western blotting analysis, the antibody reacted only weakly with the full-length TRPV1 channel protein expressed in HEK293 cultured Dells. Therefore, in order to obtain a high titer antibody against the TRPV1 E3 lap, we have established several hybridoma clones that produce monoclonal antibodies interacting with a bacterially produced fission protein containing the TRPV1 E3 loop peptide. Screening of monoclonal antibodies that specifically recognize TRPV1 protein expressed in HEK293 cells yielded several hybridoma clones. Also, it is revealed that, as expected, most of the initially established hybridomas produce monoclonal antibodies that recognize only the bacterially expressed protein. Next, we searched for the antibodies that recognize TRPV1 channel protein expressed on cell surface in non denaturing condition by live-cell ELISA. We now continue single cell cloning of a couple of hybridomas that produce monoclonal antibodies recognizing TRPV1 protein expressed on live cell surface.