人宫颈癌传代细胞Ig样蛋白的纯化分析

目的:探讨非造血系统来源的恶性肿瘤细胞内Ig样蛋白与Ig分子在抗原性及分子结构上的关系.方法:应用亲和层析的方法将HeLa MR(人宫颈癌传代细胞)细胞内Ig样蛋白进行了纯化;并以SDS-PAGE及Western Blot方法将Ig样蛋白与IgG进行比较分析.结果:SDS-PAGE结果显示Ig样蛋白既有与IgG相近的轻、重链泳带,同时又分别在相对分子质量6 600、7 000左右出现两条泳带,此两条泳带与人IgG具有高度一致的抗原性.Western Blot结果显示,Ig样蛋白与IgG相同,不但可与抗人IgG多克隆抗体、单克隆抗体反应而且可与Protein A蛋白发生反应.结论:Ig样蛋白与IgG比较在抗原性上具有明显的一致性,而在分子结构上也表现明显相似性,但二者不完全相同.

Purification and Western -blot analysis of Ig-like protein from cervical cancer cell line

Objective: To characterise the Ig like protein in non hematopietic tumor cell lines. Methods: Western blot and affinity chromatography methods were used to purify and analyze the proteins in cytoplasm of tumor cell lines. The Ig like protein was purified by affinity chromatography using staphylococal protein G and goat anti human IgG column. Results: SDS PAGE indicated that Ig like protein had not only the counterpart of the IgG heavy and light chain but also some specific bands molecular weight of 4 500, 6 600, 7 000. Western Blot analysis indicated that all the bands especially the 6 600 and 7 000 bands could react to anti human IgG polyantibodies, anti human IgG Mab and protein A. Conclusion: Protein purified by affinity chromatography showed high likeness with Ig protein. A conclusion can be drawn that Ig like protein did exist in non hematopietic tumor cell line.