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目的从培养的结肠癌细胞LOVO中纯化结肠癌相关抗原,探讨其在结肠癌患者血清中的表达。方法裂解结肠癌细胞LOVO,以抗结肠癌相关抗原单克隆抗体(mAb)4D10作为配基进行亲和层析纯化结肠癌相关抗原,SDS-PAGE电泳和Western blot进行鉴定,并用双抗夹心ELISA法检测该相关抗原在结肠癌患者及正常人血清中的表达。结果纯化所获与4D10特异结合的结肠癌相关抗原,其相对分子质量(Mr)约为65000,该抗原由Mr约为30000和35000两种亚基组成。人血清检测实验表明,该抗原在结肠癌患者血清中有较高的表达,与正常人血清相比存在显著的统计学差异(P<0.01)。结论利用mAb4D10进行免疫亲和层析,从结肠癌细胞LOVO中获得纯化的肿瘤相关抗原,该抗原有可能在临床上为结肠癌的诊断提供一定的参考价值。
Objective To purify colon cancer-associated antigen from cultured LOVO colon cancer cells and investigate its expression in the serum of colon cancer patients. Methods LOVO colon cancer cells were lysed and anti-colon cancer associated antigen mAb 4D10 was used as a ligand to purify colon cancer antigen. SDS-PAGE electrophoresis and Western blot were used to identify colon cancer cells. The relative antigen was detected in the serum of colon cancer patients and normal people. Results The relative molecular weight (Mr) of the colon cancer-associated antigen that specifically binds to 4D10 was about 65,000. The antigen consists of two subunits of Mr about 30,000 and 35,000. Human serum test showed that the antigen was highly expressed in the serum of patients with colon cancer, which was significantly different from that of normal human serum (P <0.01). Conclusion Immunoaffinity chromatography using mAb4D10 can obtain purified tumor associated antigen from colon cancer cell LOVO, which may provide some reference value for the diagnosis of colon cancer in clinic.