公开(公告)号：EP0211684A3

公开(公告)日：1988-06-08

申请号：EP86306304

申请日：1986-08-15

申请人： IMMUNEX CORPORATION

发明人： Anderson, Dirk M. ,  Cantrell, Michael A. ,  Cerretti, Douglas P. ,  Conlon, Paul J., III ,  Cosman, David J. ,  Grabstein, Kenneth H. ,  Larsen, Alf D. ,  Price, Virginia L.

IPC分类号： A61K37/02 ,  A61K35/12 ,  C12N15/00 ,  C12P21/02

CPC分类号： C07K14/535 ,  A61K38/00 ,  Y10S530/828

摘要： Macrophages and precursor monocytes are activated to exhibit tumoricidal activity by stimulation solely with granulocyte-macrophage colony stimulating factor. A patient suffering from tumors can be treated by direct administration of therapeutically effective quantities of activated granulocyte-macrophage colony stimulating factor. Homogeneous granulocyte-macrophage colony stimulating factor for use in activating macrophages and monocyte precursors is prepared by recombinant DNA techniques. The gene coding for granulocyte-macrophage colony stimulating factor is isolated and then recombinant protein product expressed in an appropriate expression system. The granulocyte-macrophage colony stimulating factor recovered from the expression system is purified to homogeneity by reverse phase high-performance liquid chromatography.

公开(公告)号：EP0215576A1

公开(公告)日：1987-03-25

申请号：EP86306303.8

申请日：1986-08-15

申请人： IMMUNEX CORPORATION

发明人： Anderson, Dirk M. ,  Baker, Paul E. ,  Cantrell, Michael A. ,  Cerretti, Douglas P. ,  Cosman, David J. ,  Gimpel, Steven D. ,  Grabstein, Kenneth H. ,  Larsen, Alf D. ,  McKereghan, Kate N.

IPC分类号： C12N15/26 ,  C12P21/02 ,  C07K13/00

CPC分类号： C07K14/55 ,  C07K2319/02 ,  C07K2319/036 ,  C07K2319/75 ,  C12N15/73

摘要： A chemically-syrtthesized oligonucleotide composing a portion of the nucleotide sequence of the human IL-2 is employed as a probe to isolate the gene coding for human IL-2. The human IL-2 gene is selected from a cDNA library prepared from RNA produced by mitogen-stimulated Jurkat cells. Double-stranded cDNA is prepared from polyadenylated RNA extracted from bovine cells thought to produce interleukin-2. Such cDNA is inserted within a plasmid vector and the recombinant plasmid employed to transform hosts. Plasmid DNA, prepared from pools of the transformed hosts, is hybridized with a probe composed of a large portion of the coding sequence of the human IL-2 gene. Pools of host cells that provide a positive signal to the human cDNA probe are identified, subdivided, and rescreened until a single positive colony is identified. Bovine plasmid cDNA is prepared from this colony, and the bIL-2 gene is sequehced. The plasmid DNA is employed to express recombinant bovine IL-2 in yeast and bacterial expression systems, with the expressad bovine IL-2 purified to homogeneity by one or more reverse phase high-performance liquid chromatography procedures.