Active site labelling of plasminogen
    1.
    发明公开
    Active site labelling of plasminogen 失效
    活性位点 - 马立氏纤溶酶原。

    公开(公告)号:EP0470494A1

    公开(公告)日:1992-02-12

    申请号:EP91112857.7

    申请日:1991-07-31

    IPC分类号: C12Q1/56 G01N33/86 C12N9/64

    摘要: A plasminogen, which has a label incorporated into an active site thereof such that said plasminogen is enzymatically and catalytically inactive upon its conversion to plasmin, and an enzymatically and catalytically inactive plasmin with a label incorporated into an active site thereof.
    A method of making a labelled plasminogen which is enzymatically and catalytically inactive upon conversion to plasmin comprising:

    a) forming a streptokinase plasminogen complex,
    b) inhibiting the plasminogen by attaching a peptide chloromethyl ketone thereto,
    c) adding a detectable molecule or marker to the peptide group derived from said peptide chloromethyl ketone thereby forming a labelled complex, and
    d) dissociating the streptokinase from the labelled plasminogen thereby forming a labelled plasminogen which is enzymatically and catalytically inactive upon conversion to plasmin.

    Preferably, the peptide chloromethyl ketone is [-(acetylthio)acetyl]-phenylalanine, phenylalanine, arginine, chloromethyl ketone and the detectable molecule or marker is a fluorophore acetamide attached to the thiol group thereof.

    摘要翻译: 一种纤溶酶原,其具有掺入其活性位点的标记,使得所述纤溶酶原在其转化为纤溶酶时是酶促和催化失活的,以及具有并入其活性位点的标记的酶和催化无活性纤溶酶。 一种制备标记的纤溶酶原的方法,其在转化为纤溶酶时是酶促和催化失活的,其包括:a)形成链激酶纤溶酶原复合物,b)通过将肽氯甲基酮连接到其上来抑制纤溶酶原,c)将可检测的分子或标记物加入到 衍生自所述肽氯甲基酮的肽基团,从而形成标记的复合物,和d)从标记的纤溶酶原解离链激酶,从而形成标记的纤溶酶原,其在转化为纤溶酶时是酶促和催化失活的。 优选地,肽氯甲基酮是[(乙酰硫基)乙酰基] - 苯丙氨酸,苯丙氨酸,精氨酸,氯甲基酮,可检测分子或标记是连接到其硫醇基上的荧光团乙酰胺。 使用在转化为纤溶酶时在酶促和催化失活的标记的纤溶酶原的测定方法包括:a)将纤溶酶原加入到样品中,和b)通过观察提供的可检测的变化来监测纤溶酶原转化为酶促和催化无活性的纤溶酶 通过标记表示催化所述转化的组分的存在。 Ë

    UROKINASE-TYPE PLASMINOGEN ACTIVATOR PROTEIN/PLASMINOGEN ACTIVATOR INHIBITOR TYPE-1 PROTEIN-SELECTED REACTION MONITORING ASSAY
    5.
    发明公开
    UROKINASE-TYPE PLASMINOGEN ACTIVATOR PROTEIN/PLASMINOGEN ACTIVATOR INHIBITOR TYPE-1 PROTEIN-SELECTED REACTION MONITORING ASSAY 审中-公开
    尿激酶型纤溶酶原激活剂,1型及含量测定监测选择的蛋白质反应的蛋白/纤溶酶原激活物抑制剂

    公开(公告)号:EP2577291A2

    公开(公告)日:2013-04-10

    申请号:EP11787438.8

    申请日:2011-05-26

    IPC分类号: G01N33/00

    摘要: Specific peptides are provided that are derived from subsequences of the urokinase-type plasminogen activator protein and the plasminogen activator inhibitor type-1 protein along with assays that can measure those peptides directly in complex protein lysate samples, including protein lysates prepared from histologicaly-processed formalin fixed tissue. The presence and amount of those peptides in samples from a subject can be associated with disease, including cancer, in a subject and provide information about the diagnostic stage/grade/status of the disease/cancer.