【摘要】 目的:制备抗人S100A4单克隆抗体(mAb),建立检测肿瘤样品中S100A4蛋白的可靠方法。方法:采用标准杂交瘤技术得到抗重组人S100A4的mAb。以ELISA、Western blot和免疫组织化学法对抗体进行鉴定。并将抗体应用于临床人乳癌和结肠癌样品的免疫组织化学检测。结果:获得了1株稳定分泌抗人S100A4蛋白mAb的杂交瘤细胞株D101。在免疫印迹实验中,该mAb较多抗特异性更好。该mAb适用于来源于人组织的样品S100A4表达的检测,在少量乳癌和结肠癌样品的检测中给出的结果与商品化多抗的结果一致。结论:该mAb是S100A4特异的,在临床应用中有更好的重复性。更多还原

【Abstract】 AIM:To prepare specific monoclonal antibody against human S100A4 protein and establish a reliable method to detect S100A4 protein in tumor specimens. METHODS: A standard hybridoma method was used to generate monoclonoal antibodies against recombinant human S100A4. ELISA, Western blot and immunohistochemistry were used to validate this antibody and the antibody is used to examine specimens of human breast and colon cancers. RESULTS: One hybridoma cell line which secreted monoclonal antibody specifically against recombinant human S100A4 protein was obtained and named as D101. The monoclonal antibody is shown to be more specific for S100A4 protein than the polyclonal antibody, at least in Western blot. The monoclonal antibody is suitable for detecting the expression of S100A4 in specimens from human tissues and gave results consistent with those of a commercially available polyclonal antibody in a small group of breast and colorectal carcinomas. CONCLUSION: The monoclonal antibody is specific for S100A4 and can be produced on a large scale; therefore it will be more reproducible for future clinical applications.更多还原