Purification of 18- and 22-kDa Forms of Basic Fibroblast Growth Factor from Rat Cells Transformed by the ras Oncogene

摘要: Normal Rat-1 fibroblasts and cells transformed by the H-ras oncogene (Rat-1-EJ) were analyzed for cell-associated growth factor activity. The two cell lines grew at same rate, but any given stage of Rat-1-EJ synthesized to four times more activity than did cells. For each line, level was five eight greater confluent densities compared sparse densities. Heparin affinity chromatography Western blot analysis demonstrated that basic fibroblast (bFGF). bFGF appeared in molecular mass forms, about 40% as an 18-kDa form which comigrated with recombinant 60% a higher doublet 22 kDa. forms biologically active could be separated on Mono S cation exchange column. Separation purification homogeneity both 22-kDa achieved combination CM-Sepharose exchange, heparin affinity-fast performance liquid chromatography, C4 reverse phase high chromatography.