Fluorescence polarization assays for high-throughput screening of neuropeptide FF receptors
Fluorescence polarization assays for high-throughput screening of neuropeptide FF receptors
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DOI：
10.1016/j.ab.2004.03.007
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发表时间：
2004-07-01
影响因子：
2.9
通讯作者：
Kim, SI
中科院分区：
文献类型：
--
作者：
Do, EU;Choi, G;Kim, SI
我们已经开发了第一个人神经肽FF2受体的荧光偏振测定在384孔微量滴定板。分析在单个孔中完成，没有转移、分离或洗涤步骤。在分析速度、位移信号幅度、精度和各种试剂的灵敏度方面，该性能适合于高通量药物筛选应用。激动剂和拮抗剂的效力的等级顺序相对于公开的放射性过滤测定法一致：DMe NPFF > NPFF >青蛙PP（蛙胰腺多肽）> PQRF酰胺> BIBF 3226。高度着色的化合物的影响是非常小的偏振信号微摩尔浓度。该方法作为一个简单而快速的替代放射性配体结合测定相关的NPFF受体的抗肥胖药物候选人。(C)2004年爱思唯尔公司All rights reserved.
We have developed the first fluorescence polarization assays of human neuropeptide FF2 receptors in 384-well microtiter plates. Assays are completed in a single well with no transfer, separation, or wash steps. The performance is suitable for high-throughput drug screening applications with regard to speed of analysis, magnitude of displaceable signal, precision, and sensitivity of various reagents. The rank order of potency of agonists and antagonists agrees well relative to the published radiometric filtration assays: DMe NPFF > NPFF > frog PP (Rana temporaria pancreatic polypeptide) > PQRFamide > BIBP 3226. The effect of highly colored compounds is very small on the polarization signal up to micromolar concentrations. The method serves as a simple and fast alternative to radioligand binding assays of antiobesity drug candidates related to NPFF receptors. (C) 2004 Elsevier Inc. All rights reserved.