METHODS AND COMPOSITIONS FOR ENHANCING DETECTION IN DETERMINATIONS EMPLOYING CLEAVABLE ELECTROPHORETIC TAG REAGENTS
申请人:Aclara BioSciences, Inc.
公开号:US20040096825A1
公开(公告)日:2004-05-20
Probe sets for the multiplexed detection of the binding of, or interaction between, one or more ligands and target antiligands are provided. Detection involves the release of identifying tags as a consequence of target recognition. The probe sets include electrophoretic tag probes or e-tag probes, comprising a detection region and a mobility-defining region called the mobility modifier, both linked to a target-binding moiety. The probes comprise interactive functionalities adjacent the cleaved portion positioned in the probes such that the interactive functionality does not form part of the e-tag reporters. Target antiligands are contacted with a set of e-tag probes and the contacted antiligands are treated with a selected cleaving agent resulting in a mixture of e-tag reporters and uncleaved and/or partially cleaved e-tag probes. The mixture is exposed to a capture agent effective to bind to uncleaved or partially cleaved e-tag probes, followed by electrophoretic separation. In a multiplexed assay, different released e-tag reporters may be separated and detected providing for target identification.
提供了一种用于多重检测一种或多种配体与目标抗配体之间的结合或相互作用的探针集。检测涉及由于目标识别而释放识别标签。探针集包括电泳标签探针或e标签探针,包括一个检测区域和一个称为迁移修饰剂的迁移定义区域,两者都链接到一个目标结合部位。探针包括与探针中定位的割裂部分相邻的交互功能,使得交互功能不构成e标签报告者的一部分。将目标抗配体与一组e标签探针接触,并用所选的切割剂处理接触的抗配体,从而产生e标签报告者和未割裂和/或部分割裂的e标签探针的混合物。将混合物暴露于有效结合到未割裂或部分割裂的e标签探针的捕获剂后进行电泳分离。在多重检测中,可以分离和检测不同的释放的e标签报告者,从而实现目标识别。