Optimization of the cost and sensitivity of receptor- and enzyme-based assays.

Optimization of the cost and sensitivity of receptor- and enzyme-based assays.
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优化基于受体和酶的检测的成本和灵敏度。

DOI:
10.1006/abio.1999.4122
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发表时间:
1999
期刊:
Analytical biochemistry.
影响因子:
--
通讯作者:
Healy,KE
Healy,KE
中科院分区:
--
文献类型:
--
作者:
Model,MA;Healy,KE

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在检测受体拮抗剂或酶抑制剂时,有三个参数通常会以可预测的定量方式影响结果:受体(酶)、标记配体(底物)和拮抗剂(抑制剂)的浓度。分析优化的通常目标是使分析检测低浓度分析物的能力最大化。另一个具有实际重要性的问题,特别是在筛选大量样品时,将是试剂成本的最小化。虽然优化受体结合试验的数学理论在很久以前就发展起来了,但所得到的公式(在配体和竞争体亲和不相等的一般情况下)并不适合实际使用。目前可用的计算程序,如Mathematica,使基于受体和酶的分析成为可能。我们使用图形方法进行分析优化,并将其应用于以下问题:(1)分析灵敏度的优化,(2)试剂成本的优化,以及(3)整个参数值范围的分析,因为数学上的最优值有时可能不切实际。计算非常简单,有时可以在几分钟内解决问题。
In detecting receptor antagonists or enzyme inhibitors, there are three parameters that often affect the outcome in a predictable quantitative manner: concentrations of the receptors (enzyme), labeled ligand (substrate), and antagonist (inhibitor). The usual goal of assay optimization is to maximize the ability of the assay to detect low concentrations of the analyte. Another question of practical importance, especially in screening of large numbers of samples, would be minimization of the reagent cost. Although the mathematical theory of optimization of the receptor binding assay was developed a long time ago, the resulting formulas (in the general case of unequal affinities of ligand and competitor) were not well suited for practical use. The current availability of computational programs, such as Mathematica, makes possible an efficient solution, both for receptor- and enzyme-based assays. We use a graphical approach to assay optimization and apply it to the following problems: (1) optimization of assay sensitivity, (2) optimization of the reagent cost, and (3) analysis of the entire range of the parameter values since the mathematically optimal values may sometimes be impractical. The computation is extremely simple and the problem can sometimes be solved in several minutes.
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