公开(公告)号：US07282339B2

公开(公告)日：2007-10-16

申请号：US10943463

申请日：2004-09-17

申请人： Joseph Beechem ,  Kyle Richard Gee ,  David Carl Hagen ,  Iain D Johnson ,  Hee-Chol Kang ,  Christina Pastula

发明人： Joseph Beechem ,  Kyle Richard Gee ,  David Carl Hagen ,  Iain D Johnson ,  Hee-Chol Kang ,  Christina Pastula

IPC分类号： G01N33/53 ,  G01N33/532 ,  G01N33/533 ,  C12Q1/42 ,  C07K1/13

CPC分类号： G01N33/542 ,  C07F9/12 ,  C07F9/65318 ,  C07F9/65522 ,  C07F9/6596 ,  C07K16/46 ,  Y10S435/968 ,  Y10S436/80 ,  Y10S530/807 ,  Y10T436/13

摘要： The present invention provides ligand-detection reagents, ligand analogs and methods for determining the presence of a ligand in a sample. The ligand-detection reagent comprises a ligand-binding antibody and a ligand analog to form an antibody-ligand analog complex wherein the ligand analog is covalently bonded to a reporter molecule. This complex may additionally comprise a labeling protein non-covalently bonded to the antibody to form a ternary complex wherein the labeling protein comprises a monovalent antibody fragment or a non-antibody protein that is covalently bonded to a label moiety. The reporter molecule is either quenched by the ligand-binding antibody or by the label moiety of the labeling protein, depending on the reporter molecule and the ligand-binding antibody, wherein the amount of quenching is directly related to the amount of ligand present in the sample. Alternatively, the ligand analog is fluorogenic wherein the ligand analog is essentially non-fluorescent in solution but when bound by the ligand-binding antibody the detectable signal increases. In this instance a decrease in signal, as opposed to the relieving of quenching, is measured for the presence of a target ligand.

摘要翻译： 本发明提供配体检测试剂，配体类似物和用于测定样品中配体存在的方法。 配体检测试剂包含配体结合抗体和配体类似物以形成抗体 - 配体类似物复合物，其中配体类似物共价键合到报告分子。 该复合物还可以包含与抗体非共价键合以形成三元复合物的标记蛋白质，其中标记蛋白质包含共价键合到标记部分的单价抗体片段或非抗体蛋白质。 取决于报道分子和配体结合抗体，报告分子被配体结合抗体或标记蛋白的标记部分淬灭，其中淬灭量与存在于 样品。 或者，配体类似物是荧光的，其中配体类似物在溶液中基本上是非荧光的，但当被配体结合抗体结合时，可检测信号增加。 在这种情况下，与目标配体的存在相比，测量了与消除淬灭相反的信号降低。