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

该项目旨在通过利用一项新发现，即称为TRPV 1的阳离子通道构成神经元细胞中传递疼痛感觉信号的最终途径之一，生产一种新的治疗性抗体，以改善持久性癌症疼痛综合征。尽管针对细菌产生的TRPV1 E3 bop产生的多克隆抗体在Western印迹分析中以高滴度识别抗原蛋白，但该抗体仅与HEK293培养的Dell中表达的全长TRPV1通道蛋白弱反应。因此，为了获得针对TRPV1 E3 lap的高滴度抗体，我们建立了几个杂交瘤克隆，其产生与细菌产生的含有TRPV1 E3环肽的分裂蛋白相互作用的单克隆抗体。筛选特异性识别HEK293细胞中表达的TRPV1蛋白的单克隆抗体产生了几个杂交瘤克隆。此外，正如预期的那样，揭示了大多数最初建立的杂交瘤产生仅识别细菌表达的蛋白质的单克隆抗体。接下来，我们通过活细胞ELISA寻找在非变性条件下识别在细胞表面表达的TRPV 1通道蛋白的抗体。我们现在继续单细胞克隆的一对夫妇的杂交瘤，产生单克隆抗体识别TRPV1蛋白表达在活细胞表面。

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.