目的:制备抗干扰素诱导的跨膜蛋白-1(interferon-induced transmembrane protein 1,IFITM1)的单克隆抗体,为检测IFITM1及进一步研究其在结肠肿瘤发生过程中的作用提供实验基础。方法:以结肠癌患者的癌组织为材料,提取总RNA,以RT-PCR扩增得到IFITM1 cDNA序列,经EcoRⅠ和HindⅢ双酶切后,克隆入pGEX-4T-3进行原核表达并纯化得IFITM1-GST;以该融合蛋白免疫BALB/c小鼠,淋巴细胞杂交瘤法制备单克隆抗体;采用ELISA、Western-blot及免疫组织化学法以制备的抗体检测结肠癌患者结肠癌组织中的IFITM1。结果:成功构建了IFITM1原核表达载体,获得了IFITM1-GST重组蛋白;制备得到了1株抗IFITM1单克隆抗体,腹水ELISA效价为1:30000,抗体亚类为IgG1,可用于ELISA、Western-blot及免疫组织化学法检测结肠癌患者结肠癌组织中的IFITM1。结论:获得了1株可用于ELISA、Western-blot及免疫组织化学法的抗IFITM1单克隆抗体2F-1,为进一步研究IFITM1在结肠肿瘤发生过程中的作用提供了实验基础。 OBJECTIVE: To prepare a monoclonal antibody against interferon-induced transmembrane protein 1 (IFITM1) and provide experimental basis for detecting IFITM1 and further studying its role in the development of colon tumor. Methods: The total RNA was extracted from the cancerous tissue of colon cancer patients. The IFITM1 cDNA sequence was amplified by RT-PCR. After digested with EcoRⅠ and HindⅢ, it was cloned into pGEX-4T-3 for prokaryotic expression and purified IFITM1 -GST. BALB / c mice were immunized with the fusion protein and monoclonal antibodies were prepared by lymphocyte hybridoma. IFITM1 in colon cancer tissues of colon cancer patients was detected by ELISA, Western-blot and immunohistochemistry. Results: The IFITM1 prokaryotic expression vector was successfully constructed and IFITM1-GST recombinant protein was obtained. One anti-IFITM1 monoclonal antibody was prepared. The ascites ELISA titer was 1: 30000 and the antibody subclass was IgG1. blot and immunohistochemistry were used to detect IFITM1 in colon cancer tissues of patients with colon cancer. CONCLUSION: An anti-IFITM1 monoclonal antibody 2F-1 that can be used for ELISA, Western-blot and immunohistochemistry was obtained, which provided the experimental basis for further study on the role of IFITM1 in the development of colonic tumors.