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    DNA encoding GM-CSF and a method of producing GM-CSF protein
    26.
    发明授权
    DNA encoding GM-CSF and a method of producing GM-CSF protein 失效
    编码GM-CSF的DNA和生产GM-CSF蛋白的方法

    公开(公告)号：US5908763A

    公开(公告)日：1999-06-01

    申请号：US287019

    申请日：1994-08-08

    申请人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    发明人： Steven C. Clark ,  Randal J. Kaufman ,  Gordon G. Wong ,  Elizabeth A. Wang

    IPC分类号： A61K38/00 ,  C07K14/535 ,  C12N15/27 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  C07K14/53 ,  C12N15/63

    CPC分类号： C12N15/85 ,  C07K14/535 ,  C12N15/70 ,  C12N15/81 ,  A61K38/00 ,  C12N2800/206 ,  C12N2830/00 ,  C12N2830/702 ,  C12N2840/105 ,  C12N2840/44

    摘要： A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises:preparing RNA from a cell that produces CSF;preparing polyadenylated messenger RNA from said RNA;preparing single stranded cDNA from said messenger RNA;converting the single stranded cDNA to double stranded cDNA;inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies;picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool;transfecting the plasmid DNA into suitable host cells for expressing CSF protein;culturing the transfected cells and assaying the supernatant for CSF activity; andselecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line. The invention also provides a method of purifying the CSF proteins and the purified proteins so produced.

    摘要翻译： 描述了制备和分离含有CSF / cDNA的转化载体的方法。 该方法包括：从产生CSF的细胞制备RNA; 从所述RNA制备聚腺苷酸化的信使RNA; 从所述信使RNA制备单链cDNA; 将单链cDNA转化为双链cDNA; 将双链cDNA插入转化载体并用所述载体转化细菌以形成菌落; 分别取200至500个菌落的池，每个池中分离出质粒DNA; 将质粒DNA转染到合适的宿主细胞中以表达CSF蛋白; 培养转染的细胞并测定上清液的CSF活性; 并选择CSF阳性池并筛选用于制备池的菌落以鉴定具有CSF活性的菌落。 还描述了编码具有CSF活性的蛋白质（即CSF / cDNA）的cDNA，用含有这种CSF / cDNA的重组载体转化的微生物或细胞系，以及通过培养表达所述CSF / cDNA来产生CSF蛋白的方法 微生物或细胞系。 本发明还提供了纯化CSF蛋白质和如此制备的纯化蛋白质的方法。

    Isolated native primate GM-CSF protein
    27.
    发明授权
    Isolated native primate GM-CSF protein 失效
    分离的天然灵长类动物GM-CSF蛋白

    公开(公告)号：US5895646A

    公开(公告)日：1999-04-20

    申请号：US344809

    申请日：1994-11-23

    申请人： Elizabeth A. Wang

    发明人： Elizabeth A. Wang

    IPC分类号： A61K38/00 ,  C07K14/535 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  C07K1/26

    CPC分类号： C07K14/535 ,  C12N15/70 ,  C12N15/81 ,  C12N15/85 ,  A61K38/00 ,  Y10S930/145

    摘要： A method for purifying CSF protein is described. The method comprises: precipitating the protein with ammonium sulfate at 80% saturation to form a pellet containing the CSF protein; resuspending the pellet in a buffered solution at a pH in the range of about 6 to about 8; applying the buffered solution containing CSF to a chromatographic column, eluting with the buffered solution containing sodium chloride and collecting the fractions having CSF activity; pooling the active fractions, applying them to a C4 reverse phase column and eluting with a 0 to 90% acetonitrile gradient to collect the active fractions. The purified CSF protein has a specific activity of at least about 1.times.10.sup.7 units per mg of protein and preferably at least about 4.times.10.sup.7 units per mg of protein when assayed using the human bone marrow assay.

    摘要翻译： 描述了用于纯化CSF蛋白的方法。 该方法包括：以饱和饱和的硫酸铵沉淀蛋白质，形成含有CSF蛋白质的沉淀物; 将沉淀物重悬于约6至约8范围内的pH的缓冲溶液中; 将含有CSF的缓冲溶液施加到色谱柱，用含有氯化钠的缓冲溶液洗脱并收集具有CSF活性的级分; 将活性级分合并，将其施加到C4反相柱上，用0至90％乙腈梯度洗脱以收集活性级分。 当使用人骨髓测定法测定时，纯化的CSF蛋白质具有至少约1×10 7单位/ mg蛋白质的比活性，优选至少约4×10 7单位/ mg蛋白质。

    Neural regeneration using human bone morphogenetic proteins
    29.
    发明授权
    Neural regeneration using human bone morphogenetic proteins 失效
    使用人骨形态发生蛋白的神经再生

    公开(公告)号：US5756457A

    公开(公告)日：1998-05-26

    申请号：US435120

    申请日：1995-05-05

    申请人： Elizabeth A. Wang ,  Josephine S. D'Alessandro ,  Dean M. Toriumi

    发明人： Elizabeth A. Wang ,  Josephine S. D'Alessandro ,  Dean M. Toriumi

    IPC分类号： A61K47/36 ,  A61K38/00 ,  A61K38/18 ,  A61L27/00 ,  A61L31/04 ,  A61P25/00 ,  A61P43/00

    CPC分类号： A61K38/1875 ,  A61L31/047 ,  A61L2430/32

    摘要： Methods and devices are disclosed for inducing growth of neural cells, and repairing neural defects in a mammal. The method comprises administering to said mammal at the site of neural defect, damage or depletion, an effective amount of a bone morphogenetic protein, either in admixture with a pharmaceutically acceptable vehicle, or adsorbed to a suitable matrix. The device comprises bone morphogenetic protein, optionally in combination with other factors, adsorbed on a suitable matrix and contained within an artificial nerve replacement vessel.

    摘要翻译： 公开了用于诱导神经细胞生长和修复哺乳动物神经缺陷的方法和装置。 该方法包括向所述哺乳动物施用神经缺陷，损伤或消耗的部位，有效量的骨形态发生蛋白，或者与药学上可接受的载体混合或吸附到合适的基质上。 该装置包括骨形态发生蛋白，任选地与其它因素组合，吸附在合适的基质上并且包含在人造神经置换容器内。