抗人肺癌单克隆抗体N-35及肺癌相关抗原N35蛋白的纯化

发布时间：2018-12-16 00:47

【摘要】： 目的利用亲和层析技术制备抗人肺癌单克隆抗体(McAb)N-35,并利用免疫亲和层析技术用自行制备的抗人肺癌单克隆抗体纯化其相应的肺癌相关抗原N35蛋白。方法培养P35杂交瘤细胞,收集培养上清,利用Protein A-Sepharose CL-4B亲和层析法纯化单克隆抗体N-35;用纯化得到的单克隆抗体与溴化氰活化的ProteinA-Sepharose CL-4B制备免疫亲和柱;再将肺腺癌GLC-82细胞的裂解液通过此免疫亲和柱,纯化肺癌相关抗原N35。结果从肺腺癌GLC-82细胞的裂解液中纯化得到分子量为80KD的N35蛋白,经SDS-PAGE分析达到了电泳纯。结论应用Protein A-Sepharose CL-4B亲和层析法纯化单克隆抗体的效率高、纯度好;应用溴化氰活化的Protein A-Sepharose CL-4B免疫亲和层析法纯化肺癌相关抗原N35也有较高的效率和纯度;纯化得到的肺癌相关抗原N35分子量为80KD,可能是一种新的肿瘤相关抗原。
[Abstract]:Objective to prepare monoclonal antibody (McAb) N-35) against human lung cancer by affinity chromatography, and to purify the corresponding lung cancer associated antigen (N35) protein with self-made monoclonal antibody against human lung cancer by immunoaffinity chromatography. Methods P35 hybridoma cells were cultured, supernatants were collected and purified by Protein A-Sepharose CL-4B affinity chromatography, and immunoaffinity columns were prepared with purified monoclonal antibodies and ProteinA-Sepharose CL-4B activated by cyanide bromide. The lung cancer associated antigen N35 was purified from the lysate of lung adenocarcinoma GLC-82 cells through the immunoaffinity column. Results N35 protein with molecular weight of 80KD was purified from the lytic fluid of lung adenocarcinoma GLC-82 cells and was purified by SDS-PAGE analysis. Conclusion the purification of monoclonal antibody by Protein A-Sepharose CL-4B affinity chromatography has high efficiency and purity, and the purification of lung cancer associated antigen N35 by Protein A-Sepharose CL-4B immunoaffinity chromatography activated by cyanide bromide has high efficiency and purity. The purified lung cancer associated antigen N35 has a molecular weight of 80 KD, which may be a new tumor associated antigen.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392.11;R734.2

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