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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。
Objective: To prepare anti-human S100A4 monoclonal antibody (mAb) and to establish a reliable method for detecting S100A4 protein in tumor samples. Methods: Anti-recombinant human S100A4 mAb was obtained using standard hybridoma technology. Antibodies were identified by ELISA, Western blot and immunohistochemistry. Antibodies were also used in immunohistochemical detection of clinical human breast and colon cancer samples. Results: A hybridoma cell line D101 with stable secreting anti-human S100A4 mAb was obtained. The mAb is more resistant to specificity in western blot experiments. The mAb is suitable for the detection of human tissue-derived sample S100A4 expression. The results given in the detection of a small number of breast and colon cancer samples are consistent with those of commercial polyclonal antibodies. Conclusion: The mAb is S100A4-specific and has better reproducibility in clinical applications.