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目的筛检出具有t EPF样活性的肿瘤样本 ,并对其进行分离纯化。方法用活性玫瑰花结抑制实验进行筛检 ,采用DEAE纤维素离子交换色谱、S SepharoseF .F离子交换色谱、ConA SepharoseCl 4B亲和色谱、Heparin SepharoseCl 6B亲和色谱等方法进行纯化 ,用SDS PAGE测定其相对分子量 ,以等电聚焦电泳法检测其等电点。结果膀胱癌、黑色素瘤、绒癌、恶性葡萄胎 4种肿瘤血清和恶性葡萄胎清宫人流血及黑色素瘤标本组织液具有t EPF活性。蛋白含量为 0 .49mg/ml,相对分子量为 134 2 4、2 32 19、382 73,等电点为 6 .5 3。结论 4种肿瘤血清和 2种肿瘤组织液中纯化获得的t EPF在相对分子量、等电点等生化指标上完全一致 ,且与胎源性EPF无本质区别
Objective To screen tumor samples with t EPF-like activity and isolate and purify them. Methods The active rosettes were screened by SDS-PAGE and purified by DEAE cellulose ion exchange chromatography, S Sepharose F F ion exchange chromatography, ConA Sepharose Cl 4B affinity chromatography and Heparin Sepharose Cl 6B affinity chromatography. SDS PAGE Its relative molecular weight, isoelectric focusing by isoelectric focusing electrophoresis. Results Bladder cancer, melanoma, choriocarcinoma, Malignant hydatidiform mole of 4 kinds of tumor serum and Malignant hydatidosis of Qing dynasties and melanoma tissue tissue fluid tEPF activity. Protein content of 0.49mg / ml, the relative molecular weight of 134 2 4,2 32 19,382 73, isoelectric point of 6,53. Conclusion The t EPF purified from 4 kinds of tumor serum and 2 kinds of tumor tissue fluid are completely consistent in biochemical indexes such as relative molecular weight and isoelectric point and have no essential difference with fetal EPF