定量放射受体分析中的离心或过滤

Centrifugation or filtration in quantitative radioreceptor assays.

作者信息

Ensing K, Feitsma K G, Bloemhof D, In 't Hout W G, de Zeeuw R A

出版信息

J Biochem Biophys Methods. 1986 Sep;13(2):85-96. doi: 10.1016/0165-022x(86)90020-5.

PMID:3772026

Abstract

In quantitative radioreceptor assays the amount of a drug present in the medium to be assayed is inversely related to the amount of receptor-bound radiolabelled ligand. Usually, separation of the bound and free fractions of radiolabelled ligand is done by filtration, in which the bound fraction can easily be collected. However, the filtration disturbs the equilibrium between bound and free fractions, which may lead to erroneous results. Because the decrease in bound radiolabelled ligand is accompanied by an increase in free labelled ligand, we decided also to measure this free fraction after separation by centrifugation and to compare these data with the filtration data. In these experiments a radioreceptor assay for anticholinergics was employed. The results indicate that both methods are compatible in precision when appropriate conditions are used whereas each method has its specific features.

摘要

在定量放射受体分析中，被测介质中存在的药物量与受体结合的放射性标记配体的量呈反比关系。通常，通过过滤来分离放射性标记配体的结合部分和游离部分，在这种情况下，结合部分很容易被收集。然而，过滤会干扰结合部分和游离部分之间的平衡，这可能导致错误的结果。由于结合的放射性标记配体的减少伴随着游离标记配体的增加，我们决定在通过离心分离后也测量这个游离部分，并将这些数据与过滤数据进行比较。在这些实验中，采用了抗胆碱能药物的放射受体分析方法。结果表明，当使用适当的条件时，两种方法在精密度上是相符的，而每种方法都有其特定的特点。