Tau蛋白外显子特异性单抗制备及初步应用

Preparation and Application of Tau Exon-Specific Monoclonal Antibodies

目前由于缺失特异性的商品化抗体,不同tau异构体在中枢神经系统疾病中,尤其在朊病毒病中的变化鲜有报道。本研究利用杂交瘤技术制备了抗tau蛋白外显子2和10的单克隆抗体,同时对制备抗体的灵敏度、特异性、免疫反应亲和力、种属反应性等方面进行了评价,随后对制备的抗体应用范围进行了摸索。结果显示,制备的四株单抗特异性高、灵敏度好,尤其可以识别小鼠脑组织中的tau蛋白。制备的四株单抗可用于间接细胞免疫荧光实验和多种感染神经细胞和感染小鼠模型脑组织的蛋白免疫印迹和免疫组织化学实验。综上,本研究制备了四株tau外显子特异性单抗,上述单抗具有较广的应用范围,将成为神经退行性疾病中tau蛋白检测的有力工具。

Changes of different tau isoforms in central nervous system diseases,especially in prion diseases have rarely been reported due to a lack of specific antibodies available commercially. In the present study,four monoclonal antibodies(mAbs) against exon 2 and exon 10 of tau proteins were prepared respectively with hybridoma technique. The prepared monoclonal antibodies were evaluated in terms of sensitivity,specificity,affinity in the immune response,and species reactivity. Subsequently,the application range of the prepared antibody was explored. Results showed that the prepared monoclonal antibodies had high specificity and sensitivity. Also,they could recognize tau protein in mouse brain specifically. Furthermore,the four prepared monoclonal antibodies could be used in cell immunofluorescence,Western Blot and immunohistochemistry experiments with various prion infected nerve cells and brain tissues from prion infected mouse models. Taken together,four tau exon-specific monoclonal antibodies were prepared and had a wide application range. These antibodies could become powerful tools for tau analyses in neurodegenerative diseases.