摘要
从肝癌病人尿液中提取和纯化出可溶性肿瘤坏死因子受体 (sTNF R)并对其生物学活性进行了检测。尿液经超滤浓缩 ,分级盐析和亲和柱层析后得到纯化的sTNF R ,经SDS PAGE显示为两条蛋白带 ,Mr 分别为 2 8× 10 4 和 5 0× 10 4 ,与国外文献报道的sTNF R1和sTNF R2 Mr 相吻合。激光扫描显示Mr2 8× 10 4 和5 0× 10 4 分别占蛋白总量的 83 44 %和 16 5 5 %。生物学研究表明 :纯化的sTNF R不仅可中和TNF和放线菌素D诱导的L92 9细胞的细胞毒性 ,并能拮抗TNF对U93
The purpose of the present investigation was to purify a urine derived soluble tumor necrosis factor receptor(sTNF R)and to characterize its activity For the purification procedure, the urine of liver cancer patients was concentrated by ultrafiltration and sTNF R purified by salt out with 40%～80% of ammonium sulphate and rhTNFα affinity column The sTNF R migrates with two protein bands on reducing sodium dodecylsulfate/polyacrylamide gel electrophoresis(SDS·PAGE) The molecular weights of two proteins are 28kD and 50kD, respectively Besides inhibiting rhTNFα induced cytotoxicity in murine L 929 fibroblasts cells in the presence of actinomycin D markedly, the sTNF R impeded also in a dose dependent manner DNA and protein synthesis in human U 937 monoblastoid leukaemic cells Therefore, sTNF R is active on both actinomycin D treated and untreated cells The sTNF R may have an important role in regulation and control of TNF activity in the cancer patients
出处
《药物生物技术》
CAS
CSCD
2000年第1期28-31,共4页
Pharmaceutical Biotechnology
关键词
可溶性
肿瘤坏死因子
受体
纯化
Soluble tumor necrosis factor receptor, Tumor necrosis factor, Purification
