抗大鼠Nogo分子单克隆抗体的制备与初步鉴定

Preparation and characterization of monoclonal antibodies against rat Nogo molecule

目的:制备抗大鼠Nogo分子单克隆抗体(mAb),为研究Nogo分子的组织分布和功能提供实验手段。方法:应用淋巴细胞杂交瘤技术制备小鼠源性抗大鼠Nogo mAb。用免疫荧光组织化学技术,对Nogo分子在大鼠中枢神经系统(CNS)的分布进行鉴定。结果:获得3株分泌抗Nogo mAb的杂交瘤细胞株。间接ELISA测定腹水效价达10-6,两株mAb为IgGl(K)亚类,另一株为IgG2b(K),在免疫荧光组织化学技术应用中获得较满意效果。结论:获得3株能特异性识别天然Nogo分子的mAb,为进一步研究Nogo分子的结构和功能奠定了基础。

AIM: To prepare monoclonal antibodies against rat Nogo molecule and study the distribution of Nogo in rat central nervous system (CMS). METHODS: Murine mAbs were prepared by hybridoma technique. The distribution of Nogo in rat CNS was identified by immunofluorescent histochemi-cal staining. RESULTS: Three hybridoma cell lines, FMU-Nogol, FMU-Nogo2 and FMU-Nogo3, secreting mAbs a-gainst rat Nogo molecule were obtained. The liters of ascitic mAbs reached to 10 -6 and the 1g subclass of FMU-Nogol, FMU-Nogo2 and FMU-Nogo3 was lgG2b(k), lgGl(k) and lgGl(k), respectively. Moreover, these mAbs could be used in immunofluoresent histochemical staining. The results showed that rat Nogo molecule could be detected in Purkinje cell layer and granual layer of rat cerebellum and on oligodendrocyte in white matter of rat spinal cord. Furthermore , these mAbs could be used in Western blot for detecting Nogo molecule present in rat spinal cord. CONCLUSION: Three mAbs against rat Nogo molecule were successfully prepared, which can provide a useful tool in research on the structure and functions of Nogo molecule in CNS.