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    • 2. 发明申请
    • SEQUENCE-SPECIFIC DETECTION OF METHYLATION IN BIOMOLECULES
    • 生物分子中甲基化的序列特异性检测
    • WO2005121361A3
    • 2006-09-28
    • PCT/EP2005006374
    • 2005-06-14
    • HAFTEN E V MAX PLANCK GES ZURWEINHOLD ELMARMEIER THOMASDUEFEL HARTMUTMARKERT-HAHN CHRISTINESCHMUCK RAINER
    • WEINHOLD ELMARMEIER THOMASDUEFEL HARTMUTMARKERT-HAHN CHRISTINESCHMUCK RAINER
    • C07H19/16C12Q1/68
    • C07H19/16C12Q1/6827C12Q2537/164C12Q2522/101C12Q2521/125
    • The present invention relates to a method for detecting sequence specific methylation in a biomolecule, comprising: (a) contacting a biomolecule with an S-adenosyl-L-methionine-dependent methyltransferase in the presence of a detectable cofactor of said methyltransferase; and (b) detecting whether the recognition sequence of said methyltransferase has been modified with the cofactor or a derivative thereof, wherein modification of the recognition sequence of said methyltransferase is indicative of an absence of methylation at said recognition sequence. The present invention also relates to a cofactor which is specific for S-adenosyl-L-methionine-dependent methyltransferases, wherein said cofactor is an N-adenosylaziridine derivative with a reporter group attached to the 6 or 7 position of the adenine ring or attached to the aziridine ring. Moreover, the present invention relates to a complex of the cofactor of the present invention and a methyltransferase which normally uses S-adenosyl-L-methionine (AdoMet) as a cofactor. In addition, the present invention relates to a diagnostic composition comprising the cofactor of the present invention or the complex of the present invention. Finally, the present invention relates to the use of the cofactor of the present invention or the complex of the present invention for detecting sequence-specific methylation in DNA molecules.
    • 本发明涉及用于检测生物分子中序列特异性甲基化的方法,其包括:(a)在所述甲基转移酶的可检测辅因子存在下使生物分子与S-腺苷-L-甲硫氨酸依赖性甲基转移酶接触; 和(b)检测所述甲基转移酶的识别序列是否已经用辅因子或其衍生物修饰,其中所述甲基转移酶的识别序列的修饰指示在所述识别序列处不存在甲基化。 本发明还涉及对S-腺苷-L-甲硫氨酸依赖性甲基转移酶特异性的辅因子,其中所述辅因子是N-腺苷氮丙啶衍生物,其报告基团连接到腺嘌呤环的6或7位或附着于 氮丙啶环。 此外,本发明涉及本发明的辅因子与通常使用S-腺苷-L-甲硫氨酸(AdoMet)作为辅因子的甲基转移酶的复合物。 此外,本发明涉及包含本发明的辅因子或本发明的复合物的诊断组合物。 最后,本发明涉及本发明的辅因子或本发明的复合物用于检测DNA分子中的序列特异性甲基化的用途。
    • 3. 发明申请
    • METHOD FOR HIGH-THROUGHPUT SCREENING OF TEST MOLECULES BINDING TO TARGET MOLECULES
    • 测试分子与目标分子结合的高通量筛选方法
    • WO2006015797A3
    • 2006-05-11
    • PCT/EP2005008464
    • 2005-08-04
    • HAFTEN E V MAX PLANCK GES ZURSAUER SASCHABUESSOW KONRADLEHRACH HANS
    • SAUER SASCHABUESSOW KONRADLEHRACH HANS
    • G01N33/537G01N33/538
    • G01N33/538G01N2500/04
    • This invention relates to a method for identifying one or more molecules binding to a target molecule in a mixture of one or more test molecules, each test molecule having at least one known characteristic property, wherein the method comprises the following steps: (a) bringing said one or more test molecules into contact with said target molecule under conditions allowing formation of a complex comprising said target molecule and the molecule(s) binding to said target molecule; (b) separating said complex or said target molecule from non-binding test molecule(s); (c) detecting said non-binding test molecule(s), if any, via their known characteristic property/ies; and (d) identifying said molecule(s) binding to said target molecule, if any, being the test molecule(s) not detected in step (c). In a preferred embodiment, detection is effected by mass spectrometry. In a further preferred embodiment, separation is effected by size exclusion chromatography.
    • 本发明涉及用于鉴别在一种或多种测试分子的混合物中结合靶分子的一种或多种分子的方法,每种测试分子具有至少一种已知的特征性质,其中该方法包括以下步骤:(a)将 所述一个或多个测试分子在允许形成包含所述靶分子和与所述靶分子结合的分子的复合物的条件下与所述靶分子接触; (b)将所述复合物或所述靶分子与非结合测试分子分离; (c)通过其已知特征性质检测所述非结合测试分子(如果有的话); 和(d)鉴定与步骤(c)中未检测到的测试分子结合的所述目标分子(如果有的话)的所述分子。 在优选的实施方案中,通过质谱法进行检测。 在另一优选实施方案中,通过尺寸排阻色谱法进行分离。