公开(公告)号：US20080311674A1

公开(公告)日：2008-12-18

申请号：US11837427

申请日：2007-08-10

申请人： Sharat Singh ,  Hossein Salimi-Moosavi ,  Syed Hasan Tahir ,  Gerald J. Wallweber ,  Hrair Kirakossian ,  Tracey J. Matray ,  Vincent S. Hernandez

发明人： Sharat Singh ,  Hossein Salimi-Moosavi ,  Syed Hasan Tahir ,  Gerald J. Wallweber ,  Hrair Kirakossian ,  Tracey J. Matray ,  Vincent S. Hernandez

IPC分类号： G01N27/26 ,  G01N33/53

CPC分类号： G01N33/6842 ,  C07K16/00 ,  C07K2317/40 ,  G01N33/582 ,  G01N33/6803

摘要： Methods, compositions and kits are disclosed for determining one or more target polypeptides in a sample where the target polypeptides have undergone a post-translational modification. A mixture comprising the sample and a first reagent comprising a cleavage-inducing moiety and a first binding agent for a binding site on a target polypeptide is subjected to conditions under which binding of respective binding moieties occurs. The binding site is the result of post-translational modification activity involving the target polypeptide. The method may be employed to determine the target polypeptide itself. In another embodiment the presence and/or amount of the target polypeptide is related to the presence and/or amount and/or activity of an agent such as an enzyme involved in the post-translational modification of the target polypeptide. The interaction between the first binding agent and the binding site brings the cleavage-inducing moiety into close proximity to a cleavable moiety, which is associated with the polypeptide and is susceptible to cleavage only when in proximity to the cleavage-inducing moiety. In this way, an electrophoretic tag for each of the polypeptides may be released. Released electrophoretic tags are separated and the presence and/or amount of the target polypeptides are determined based on the corresponding electrophoretic tags.

摘要翻译： 公开了用于确定样品中一种或多种靶多肽已经经历了翻译后修饰的方法，组合物和试剂盒。 包含样品和包含切割诱导部分的第一试剂和用于靶多肽上的结合位点的第一结合剂的混合物经受各自结合部分的结合发生的条件。 结合位点是涉及靶多肽的翻译后修饰活性的结果。 该方法可用于确定靶多肽本身。 在另一个实施方案中，靶多肽的存在和/或量与试剂例如涉及靶多肽的翻译后修饰的酶的存在和/或量和/或活性有关。 第一结合剂和结合位点之间的相互作用使切割诱导部分紧密接近可切割部分，其可与多肽相关，并且仅在接近切割诱导部分时易于切割。 以这种方式，可以释放每个多肽的电泳标签。 分离释放的电泳标签，并且基于相应的电泳标签确定靶多肽的存在和/或量。