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目的纯化并分析肺癌相关抗原的有关特性。方法采用商品化麦芽凝聚素 Wheatgerm L ectin- Sepharose- 6 MB层析柱和自制抗人肺癌单克隆抗体 N- 35 - Sepharose- 4B层析柱纯化肿瘤相关抗原 ,比较纯化结果 ;用免疫印技术测定肺癌相关抗原在肺癌、肝癌、宫颈癌、乳腺癌细胞及正常人心脏及肺组织的存在及分布情况 ;用差速离心技术分析肺癌相关抗原在亚细胞结构的布状况 ;用免疫荧光技术探测肺癌相关抗原在肿瘤细胞有丝分裂过程中的染色体结构定位。结果经麦芽凝聚素纯化的肺癌相关抗原 N35以不同分子量形式存在于肺癌、肝癌、宫颈癌及乳腺癌细胞的蛋白组分中 ,在正常人心脏及肺组织中无表达 ;在亚细胞结构中主要分布于胞核 ,线粒体次之 ,胞膜上分布最少 ,主要不以膜蛋白或跨膜蛋白的形式存在 ,并定位于肿瘤细胞有丝分裂中期 (methaphase)的中心粒 (centriole)结构上。结论肺癌相关抗原 N35是一种重要的肿瘤相关抗原 ,其功能很可能与肿瘤细胞无限制增殖活动有关。麦芽凝集素亲和层析法是纯化肺癌相关蛋白 N35简便有效的途径。
Objective To purify and analyze the relevant characteristics of lung cancer related antigens. Methods Tumor - associated antigens were purified by commercial Wheatgerm Lectin - Sepharose - 6 MB column and self - made anti - human lung cancer monoclonal antibody N - 35 - Sepharose - 4B column. The purification results were compared with each other. Lung cancer, cervical cancer, breast cancer cells and normal human heart and lung tissue of the presence and distribution; using differential centrifugation analysis of lung cancer-related antigen in the sub-cellular structure of cloth; using immunofluorescence detection of lung cancer Chromosomal structural localization of related antigens during mitosis in tumor cells. Results The N35, a lung cancer-associated antigen purified by malnaglutination, existed in the protein components of lung cancer, liver cancer, cervical cancer and breast cancer cells with different molecular weight and had no expression in the normal human heart and lung tissues. In the subcellular structure, Distributed in the nucleus, followed by the mitochondria, with the least distribution on the membrane, not mainly in the form of membrane proteins or transmembrane proteins, and localized on the centriole structure of the metaphase of tumor cells. Conclusion The lung cancer-associated antigen N35 is an important tumor-associated antigen and its function is likely to be related to the unlimited proliferative activity of tumor cells. Malt agglutinin affinity chromatography is a simple and effective way to purify lung cancer related protein N35.