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目的：分析和确定N35肺癌相关蛋白的有关特性及其潜在的临床应用价值。方法：利用抗人肺癌单克隆抗体N35作为免疫探针，经免疫沉淀和免疫印迹法测定其相关抗原在肺癌细胞系GLC82、宫颈癌细胞系Hela、肝癌细胞系HepG2、乳腺癌细胞系PMC、正常人心脏及肺组织中的存在及分布情况；用N聚糖酶酶解方法确定肿瘤相关蛋白N35与糖蛋白分子的关系；采用差速离心技术分离出肺腺癌细胞系GLC82亚细胞结构中的胞膜、胞核及线粒体成分，分别取样经免疫印迹法测定肿瘤相关蛋白N35在亚细胞结构中的分布状况并与免疫组化结果相印证；用免疫荧光技术探测肿瘤相关蛋白N35在肿瘤细胞有丝分裂过程中的染色体结构定位；用多种免疫亲和层析技术尝试获得大量纯化肿瘤相关蛋白N35的有效方法。结果：肿瘤相关蛋白N35是一种糖蛋白分子，不存在于正常人心脏及肺组织蛋白组分中，而以不同分子量形式分布于GLC82、Hela、HepG2和PMC细胞蛋白中；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，似不以膜蛋白或跨膜蛋白的形式存在；在肿瘤细胞有丝分裂进入S至G2期时明确地定位于中心粒（centriole）结构上，强烈提示其功能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大量纯化肿瘤相关蛋白N35最有效的途径。结?
Objective: To analyze and determine the relevant characteristics of N35 lung cancer related protein and its potential clinical value. Methods: The anti-human lung cancer monoclonal antibody N35 was used as an immunoprecipitation probe. The relative antigens were detected by immunoprecipitation and immunoblotting in lung cancer cell line GLC82, cervical cancer cell line Hela, liver cancer cell line HepG2, breast cancer cell line PMC, Human heart and lung tissue; the relationship between tumor-associated protein N35 and glycoprotein molecules was determined by N-glycan enzymolysis method; and the differential centrifugation was used to isolate the subcellular structure of lung adenocarcinoma cell line GLC82 Cell membrane, nucleus and mitochondria. The distribution of tumor-associated protein N35 in sub-cellular structure was determined by immunoblotting and confirmed with immunohistochemistry. Immunofluorescence was used to detect the expression of tumor-associated protein N35 in mitosis Chromosome structure of the process of positioning; using a variety of immunoaffinity chromatography techniques to try to obtain a large number of purification of tumor-associated protein N35 an effective method. Results: The tumor-associated protein N35 is a glycoprotein which is not found in the normal human heart and lung tissue but distributed in GLC82, Hela, HepG2 and PMC cell proteins in different molecular weights. In the subcellular structure Mainly distributed in the nucleus, followed by the mitochondria, with the least distribution on the membrane, which does not seem to exist as membrane proteins or transmembrane proteins; it is specifically localized on the centriole structure when the tumor cells enter mitosis S to G2 , Strongly suggesting that its function may be related to the unrestricted proliferation of tumor cells. Maltogenic lectin affinity chromatography is the most effective way to purify the tumor-associated protein N35 in large quantities. Knot?