Development of a lanthanide-based assay for detection of receptor-ligand interactions at the δ-opioid receptor
Development of a lanthanide-based assay for detection of receptor-ligand interactions at the δ-opioid receptor
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DOI：
10.1016/j.ab.2005.05.040
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发表时间：
2005-08-15
影响因子：
2.9
通讯作者：
Gillies, RJ
中科院分区：
文献类型：
--
作者：
Handl, HL;Vagner, J;Gillies, RJ
镧系元素为基础的配体-受体相互作用的测定提供了一个有吸引力的替代传统的放射性标记的测定灵敏度，吞吐量和生物危害。我们设计并测试了五种肽配体的δ-阿片受体，用含铕（Eu）的螯合物修饰。测试这些标记的配体的结合亲和力，并与未标记的亲本配体进行比较。Eu-二亚乙基三胺五乙酸（DTPA）-[D-Pen（2），L-Cys（5）]脑啡肽（DPLCE）配体与过表达人δ-阿片受体的中国仓鼠卵巢（CHO）细胞结合，亲和力与未标记配体相似。该配体用于竞争性结合试验，结果与使用传统放射性标记结合试验获得的结果相当。这些基于镧系元素的测定提供了具有更高通量的上级结果，并且消除了对放射性废物处置的需要;因此，它们适合于配体文库的高通量筛选。(C)2005年爱思唯尔公司All rights reserved.
A lanthanide-based assay for ligand-receptor interactions provides an attractive alternative to the traditional radiolabeled determinations in terms of sensitivity, throughput, and biohazards. We designed and tested five peptide ligands for the delta-opioid receptor that were modified with a europium (Eu)-containing chelate. These labeled ligands were tested for their binding affinities and compared with the unlabeled parental ligands. The Eu-diethylenetriaminepentaacetic acid (DTPA)-[D-Pen(2), L-Cys(5)] enkephalin (DPLCE) ligand bound to Chinese hamster ovary (CHO) cells overexpressing the human delta-opioid receptor with affinity similar to the unlabeled ligand. This ligand was used in competitive binding assays with results comparable to those obtained using the traditional radiolabeled binding assays. These lanthanide-based assays provide superior results with higher throughput and eliminate the need for radioactive waste disposal; hence, they are appropriate for high-throughput screening of ligand libraries. (C) 2005 Elsevier Inc. All rights reserved.