抗人尿道高分化鳞癌单克隆抗体的研制

Development of Monoclonal Antibodies of Highly Differentiated Squamous Cell Carcinoma in Epithelioid Tumour Tissue

用本室建立的人尿道高分化鳞状上皮癌细胞株(Hus-98)免疫Balb/C小鼠,通过细胞杂交技术及ELISA方法进行筛选,SP免疫组织化学方法进行鉴定,获得10株能稳定分泌、选择性强、效价高的杂交瘤细胞株;同时从乳腺癌组织中提出肿瘤相关抗原行免疫印记实验(Western-blot)。结果6株经4次克隆能稳定分泌抗体Ig,免疫组化显示这些抗体是针对来源于上皮的鳞癌、腺癌细胞膜和细胞浆的,而癌旁正常组织及胎儿组织呈阴性反应;通过western-blot结果显示肿瘤相关抗原分子量为46.414kb。

To found hybridoma cell of squamous-cell-carcinoma-related antigen (SCC Ag) and identify the specificity of these monoclonal antibodies and ascertain the molecular weight of the corresponding protein of the monoclonal antibody of squamous cell carcinoma in urethra by western blot at the same time, the highly differentiated squamous cell carcinoma in epithelioid(Hus-98) was used to immunize the Balb/C mouse, and was screened by technology of hybridization and ELISA. Identification was done by the method of SP. We got ten hybridoma cells with steady excretion and high selectivity and high titer. Six of ten cells could excrete antibody Ig by four-time clone. The result of immunohistochemistry showed that tumour tissues presented positive dye in the cell membrane and cell plasm in squamous cell were carcinoma and gland carcinoma, but normal tissues presented negative dye.And the result of Western-blot showed that the SCC Ag was 46.414kb.