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    • 9. 发明申请
    • USE OF 14-3-3 PROTEINS AND A METHOD FOR DETERMINING THE SAME IN THE FLUIDS OR TISSUES OF ORGANISMS
    • 14-3-3蛋白用途和液体或组织生物体法的测定的
    • WO02099434A2
    • 2002-12-12
    • PCT/EP0206139
    • 2002-06-04
    • MUELLER WERNER E GSCHROEDER HEINZ C
    • MUELLER WERNER E GSCHROEDER HEINZ C
    • G01N33/68
    • G01N33/6896G01N2800/2828
    • The aim of the invention is to provide an identification method and a quantification method for 14-3-3 proteins and their isoforms for the early diagnosis of TSE diseases, which allows the diagnosis to be carried out in living organisms. A further aim is to identify a contamination of the sample by the parallel determination of a second antigen. This is achieved by the use of the biochemical characteristics of a representative of the 14-3-3 protein family, which bind to specific amino acid motives such as X(n)-XSX(n)SXXSX-X(n) or to the motive RSXpSXP within peptides or proteins. Modified solid phases, such as for example microtitre plates, are used to determine one or several isoforms or the entire 14-3-3 protein(s) and for specific binding. Said plates are coated with an artificial or natural peptide, which contains a binding motive for 14-3-3 proteins, for example a chemically synthesised peptide with the motive CAALPKINRSApSEPSLHR. After addition of the extracts or bodily fluids to be analysed, the formed peptide 14-3-3 protein complexes are detected and quantified using tagged antibodies. The inventive application of the 14-3-3 protein family and/or individual isoforms of the 14-3-3 proteins can be used as an effect monitor or biomonitor in aquatic invertebrates of environmental influences such as polychlorinated biphenyls (PCBs), (Xeno)oestrogens or other influences. The method can also be used for the early diagnosis of TSE diseases, such as Creutzfeldt-Jakob disease (CJD) and its new form (variant) in young people (vCJD) and Bovine Spongiform Encephalopathy (BSE) or similar diseases. The invention thus provides a diagnosis tag (surrogate tag), which can be used in the living organism as a screening, confirmation or sole tag.
    • 要进行的本发明中,14-3-3蛋白或它们对TSE疾病的早期诊断亚型的检测和定量方法的方法的目的可用的,其允许执行对生物体的诊断。 此外,它是由平行测定的第二抗原的检测样本的污染的目的。 根据本发明的该目的是通过使用14-3-3蛋白家族,代表的生化特性的实现,其在特定的氨基酸基序,如X(n)的-XSX(n)的SXXSX-X(n)或肽内的主题RSXpSXP或 蛋白质结合。 对于一种或多种同种型的确定或所有14-3-3蛋白(S)和特异性修饰固相结合,例如,例如,微量滴定板,涂有具有用于14-3的结合基序的合成的或天然的肽的 -3含蛋白,例如 化学合成的肽具有基序CAALPKINRSApSEPSLHR。 此外,提取物的要被测试或之后体液,所形成的肽14-3-3蛋白复合物的检测和定量是使用标记的抗体进行检测。 本发明的应用他14-3-3蛋白家族和/或14-3-3蛋白的同种型个体可以用作使用效果监视器或生物监测水生无脊椎动物对环境的影响,例如多氯联苯(PCB),(异种)的雌激素或其它。 此外,也可以在年轻人(贾氏)和牛海绵状脑病(BSE)或类似的疾病用于TSE疾病如克雅氏病(CJD)和它的新的形式(变体)的早期诊断方法。 这提供了可以用作在活生物体认证Screenings-或单个标记物的诊断用标记物(替代标记)可用。