Non-isotopic receptor assay for benzodiazepines using a biotin-labeled ligand and biotin-immobilized microtiter plate.
Non-isotopic receptor assay for benzodiazepines using a biotin-labeled ligand and biotin-immobilized microtiter plate.
复制标题
使用生物素标记配体和生物素固定微量滴定板对苯二氮卓类药物进行非同位素受体测定。
DOI：
10.1016/0021-9673(92)80143-i
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Rechnitz,GA
中科院分区：
文献类型：
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作者：
Tanaka,S;Takeuchi,T;Rechnitz,GA
采用生物素标记配体生物素- 1012s，建立了苯二氮卓类药物的非同位素受体测定方法。采用简单吸附法将生物素化的牛血清白蛋白（biotin-BSA）固定在微滴板孔壁上。由于其与生物素有较强的相互作用，可以从溶液中提取出亲和素过氧化物酶偶联物。然后通过测量酶活性来确定在壁上摄取的亲和素过氧化物酶的量。壁上固定生物素与游离生物素对亲和素的竞争为固相亲和素-生物素结合试验提供了基础。该结合法不仅可以灵敏地测定生物素，还可以测定生物素- 1012s。由于1012S是一种对苯二氮卓类受体具有高亲和力的配体，因此生物素-1012S可以用作非同位素受体检测的探针配体。基于生物素- 1012s和各种苯二氮卓类药物对受体结合位点的竞争，证明了一种非同位素受体测定方法。
A non-isotopic receptor assay for benzodiazepine drugs was developed using a biotin-labeled ligand, biotin-1012S. Biotinylated bovine serum albumin (biotin-BSA) was immobilized onto the wall of microtiter plate wells by simple adsorption. Avidin peroxidase conjugate could be extracted from solution owing to its strong interaction with biotin. The amount of avidin peroxidase taken up on the wall was then determined by measuring the enzyme activity. The competition between immobilized biotin on the wall and free biotin for avidin provided the basis for a solid-phase avidin-biotin binding assay. By this binding assay, not only biotin but also biotin-1012S could be measured sensitively. Because 1012S is a ligand with high affinity to benzodiazepine receptors, biotin-1012S could be utilized as a probe ligand for a non-isotopic receptor assay. Based upon the competition between biotin-1012S and various benzodiazepine drugs for the receptor binding sites, a non-isotopic receptor assay was demonstrated.