抗人类癌抗原单克隆抗体的制备与肝组织中的鉴定

Preparation and identification of anti-HCA monoclonal antibody

目的运用杂交瘤技术制备人类癌抗原(Human carcinoma antigen,HCA)的IgG单克隆抗体。方法用现有的HCA-IgM抗体HAE3分别从PC3、PCaT及LCaT总蛋白中分离纯化得到三组糖蛋白复合物组分作为抗原分别免疫Balb/c小鼠,获得鼠抗人HCA抗体.并对这些抗体进行ELISA检测效价、western-blot以及免疫组化等分析,筛选合适的抗体进行下一部分研究。结果共制备出62株高亲和力、高特异性并且针对不同抗原结合位点的鼠抗人HCA单克隆抗体。通过在肝组织上的鉴定,发现这些抗体无论效价、western-blot以及免疫组化都适合进行进一步研究。结论我们利用天然HCA抗原制备了高亲和力,高特异性并且针对不同抗原结合位点的单克隆抗体,为我们进一步研究其结构及功能提供了有力的特异的工具。

Objective To prepare murine mAbs against human HCA by hybridoma technique. Methods We isolated HCA from prostatic carcinoma cells （ PC3 ）, prostatic carcinoma tissues （PCaT） and lung cancer tissues （LCaT） with immunoaffinity chro- matography. In order to obtain antibodies with high affinity and specificity against HCA, we used all of these three parts of HCA to im- munize Balb/c mice and generated hybridoma cell strains secreting murine mAbs against human HCA by hybridoma technique. The an- tibodies against HCA were selected by Western blot, ELISA and immunohistochemistry. Result 62 hybridoma cell lines stably secre- ting specific mAbs against human HCA were established. The murine mAb against HCA would be efficient affinity reagent for the fur- ther functional studies of HCA on liver tissues. Discussion The murine mAb against HCA would be efficient affinity reagent for the further functional studies of HCA.