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    • 4. 发明授权
    • Processes for the purification of human recombinant decorin and the
detection of guanidinium ions
    • 人重组核心蛋白聚糖纯化方法和胍离子的检测
    • US5567807A
    • 1996-10-22
    • US272919
    • 1994-07-08
    • William S. CraigJohn R. HarperPaul J. KostelJonathan R. ParkerThomas S. Vedvick
    • William S. CraigJohn R. HarperPaul J. KostelJonathan R. ParkerThomas S. Vedvick
    • G01N30/02A61K38/00A61P35/00A61P43/00B01D41/04C07K14/00C07K14/47C07K14/78C12N15/09C12Q1/00G01N30/06G01N30/26G01N30/46G01N30/88G01N30/96G01N33/53G01N33/68C07K1/20
    • G01N33/5308C07K14/4725G01N33/68Y10T436/173845
    • The present invention is directed to a process for the production of substantially pure human recombinant decorin which involves the combination of three separate stages characterized by contacting decorin-containing cell culture medium with (1) a first strong anionic exchange resin; then with (2) a hydrophobic interactive chromatographic resin; and finally with (3) a second strong anionic exchange resin. By using a combination of steps, and certain reagents, in particular a 2.4 to 3 molar GuHCl solution to elute decorin from a hydrophobic interactive column, the process of this invention provides a more convenient and reproducible process for purifying human recombinant decorin. The invention also provides a process for detecting the presence of guanidinium ions in a sample solution. The detection process involves contacting a sample solution suspected of containing guanidinium ions with a cation exchange resin and eluting the guanidinium ions present in tire sample solution with an aqueous buffer solution having a pH of about 1.5 to about 2. This is followed by contacting the eluant with a cation suppressor columns and simultaneously flowing a suppressor regenerate solution in the opposite direction on the opposite side of the permeable membrane of the column, and finally, detecting the presence of guanidinium ions in the eluant from the ion exchange column which was contacted with the suppressor column by use of a conductivity detector.
    • 本发明涉及生产基本上纯的人重组核心蛋白聚糖的方法,其涉及三个独立阶段的组合,其特征在于使含有核蛋白聚糖的细胞培养基与(1)第一强阴离子交换树脂接触; 然后用(2)疏水交互色谱树脂; 最后用(3)第二强阴离子交换树脂。 通过使用步骤的组合和某些试剂,特别是2.4至3摩尔的GuHCl溶液从疏水相互作用的色谱柱洗脱核心蛋白聚糖,本发明的方法提供了一种更方便和可重现的纯化人重组核心蛋白聚糖的方法。 本发明还提供了检测样品溶液中胍离子的存在的方法。 检测过程包括将怀疑含有胍离子的样品溶液与阳离子交换树脂接触,并用pH为约1.5至约2的pH缓冲溶液洗脱存在于轮胎样品溶液中的胍离子。然后将洗脱液 用阳离子抑制柱,同时在柱的可渗透膜的相对侧沿相反方向流动抑制剂再生溶液,最后,检测离离子交换柱的洗脱液中胍基离子的存在,该离子交换柱与 抑制柱通过使用电导率检测器。