肿瘤内皮细胞标志物8单抗的制备及组织分布

目的　制备特异性抗人肿瘤内皮细胞标志物 8(TEM8)单克隆抗体 ;检测TEM 8蛋白在部分肿瘤和正常组织中的分布。方法　原核表达含TEM 8膜外段多肽的融合蛋白 ,免疫Balb/c雌性小鼠后取效价较高的小鼠脾细胞与SP2 / 0骨髓瘤细胞进行细胞融合 ,以ELISA、Westernblot筛选、鉴定分泌特异性抗TEM8的单克隆杂交瘤细胞株 ;ABC免疫组织化学方法检测TEM 8蛋白表达产物在结肠癌、前列腺癌、乳腺癌、结肠癌旁组织等组织中表达。结果　共获得 5株稳定分泌TEM 8抗体的单克隆杂交瘤细胞株 ;TEM 8在多种组织细胞表达 :结肠癌新生血管 (4 / 2 3) ,结肠癌癌性腺上皮 (8/ 2 3) ,结肠癌旁组织腺上皮 (6 / 17) ,结肠组织平滑肌 (17/ 37) ,前列腺癌腺上皮 (1/ 17)。结论　成功制备抗人TEM 8单克隆抗体 ;TEM8在蛋白质水平并非肿瘤血管特异性标志物

Preparation of Anti-Human Tumor Endothelial Marker 8 Specific Monoclonal Antibody and Distribution of TEM8 Protein in Tissues

Purpose To prepare anti-human tumor endothelial marker 8 monoclonal antibody,and observe the distribution of TEM8 protein in some normal and malignant human tissues. Methods The TEM8 protein expressed by prokaryotic vector was used to immunize female Balb/c mice. After cell fusion,hybridoma cells were screened for specific antibody production by ELISA and Western blot. The expression of TEM8 was detected with immunohistochemistry in colorectal carcinoma, prostate cancer, breast cancer,the normal colorectal tissue around carcinoma. Results Five hybridoma cell lines producing the specific monoclonal antibody against TEM8 have been obtained. Besides the endothelial cells of colorectal carcinoma,TEM8 protein was also detected in other cells such as malignant and normal epithelium of colorectum and prostate, smooth muscle cells of colorectum. Conclusions Anti-TEM8 specific monoclonal antibody was successfully developed. TEM8 protein is not specifically distributed in tumor vasculture.