Suppr超能文献

基于滴定法的筛选在高通量分析快速先导测试中的应用

Application of Titration-Based Screening for the Rapid Pilot Testing of High-Throughput Assays.

作者信息

Zhang Ji-Hu, Kang Zhao B, Ardayfio Ophelia, Ho Pei-i, Smith Thomas, Wallace Iain, Bowes Scott, Hill W Adam, Auld Douglas S

机构信息

Center for Proteomic Chemistry, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.

Center for Proteomic Chemistry, Novartis Institutes for Biomedical Research, Cambridge, MA, USA

出版信息

J Biomol Screen. 2014 Jun;19(5):651-60. doi: 10.1177/1087057113512151. Epub 2013 Nov 18.

Abstract

Pilot testing of an assay intended for high-throughput screening (HTS) with small compound sets is a necessary but often time-consuming step in the validation of an assay protocol. When the initial testing concentration is less than optimal, this can involve iterative testing at different concentrations to further evaluate the pilot outcome, which can be even more time-consuming. Quantitative HTS (qHTS) enables flexible and rapid collection of assay performance statistics, hits at different concentrations, and concentration-response curves in a single experiment. Here we describe the qHTS process for pilot testing in which eight-point concentration-response curves are produced using an interplate asymmetric dilution protocol in which the first four concentrations are used to represent the range of typical HTS screening concentrations and the last four concentrations are added for robust curve fitting to determine potency/efficacy values. We also describe how these data can be analyzed to predict the frequency of false-positives, false-negatives, hit rates, and confirmation rates for the HTS process as a function of screening concentration. By taking into account the compound pharmacology, this pilot-testing paradigm enables rapid assessment of the assay performance and choosing the optimal concentration for the large-scale HTS in one experiment.

摘要

使用小型化合物集对用于高通量筛选(HTS)的分析方法进行预试验是分析方法验证过程中必要但通常耗时的一步。当初始测试浓度不理想时,这可能需要在不同浓度下进行反复测试,以进一步评估预试验结果,这可能会更加耗时。定量高通量筛选(qHTS)能够在单个实验中灵活、快速地收集分析性能统计数据、不同浓度下的活性化合物以及浓度-响应曲线。在此,我们描述了预试验的qHTS过程,其中使用板间不对称稀释方案生成八点浓度-响应曲线,前四个浓度用于代表典型HTS筛选浓度范围,后四个浓度用于进行稳健的曲线拟合以确定效价/效能值。我们还描述了如何分析这些数据,以预测作为筛选浓度函数的HTS过程中的假阳性、假阴性、命中率和确认率。通过考虑化合物药理学,这种预试验范式能够在一个实验中快速评估分析性能并选择大规模HTS的最佳浓度。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验