抗ARL-1单克隆抗体的制备及初步应用

目的: 制备抗醛糖还原酶相似蛋白(aldosereductaselikeprotein, ARL- 1)的单克隆抗体 (mAb)并进行初步应用。方法: 用纯化的ARL- GST蛋白免疫BALB/c小鼠, 采用杂交瘤技术制备mAb。间接ELISA法、Westernblot及免疫组织化学染色法, 对mAb进行鉴定及初步应用。结果: 获得 1株可稳定分泌抗ARL -1蛋白mAb的杂交瘤细胞系。mAb的Ig亚类为IgG2b, 轻链属于κ型。腹水mAb的效价为: 1∶4. 096×105。Westernblot的结果表明, ARL 1蛋白在被检测的肝癌组织中呈高表达, 而在相应的癌旁组织中不表达。免疫组织化学染色的结果表明, ARL -1蛋白在肝癌组织中呈高表达且主要定位在胞质内。结论: 成功地制备 1株抗ARL -1蛋白的mAb。初步应用的结果表明, ARL- 1蛋白在肝癌组织中呈高表达, 为进一步研究ARL -1蛋白的功能, 以及其与肝癌的确切关系奠定了基础。

Preparation and characterization of monoclonal antibody against ARL-1 protein

AIM: To prepare monoclonal antibodies against aldose reductase-like(ARL-1)protein. METHODS: Purified ARL-1 protein (ARL-GST) was used to immunize BALB/c mice, and mAbs were prepared by hybridoma technique. The mAbs against ARL-1 were detected by ELISA, Western blot and immunohistochemical staining respectively. RESULTS: One clone of hybridoma secreting specific mAb against ARL-1 was obtained. The Ig subclass of mAb was of IgG2b and the titre of the antibody in ascites was 1:4.096 x 10(5). Highexpression of ARL-1 protein in liver cancer was detected. CONCLUSION: The specific mAb against ARL-1 lays the foundation for investigation of functions of ARL-1 protein and relationship between ARL-1 protein and liver cancer.