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Validation of Low-Volume 1536-Well Assay-Ready Compound Plates

机译:小容量1536孔分析就绪复合板的验证

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摘要

Assay-ready compound plates (ARPs) are sealed assay plates that contain DMSO solutions of screening compounds predispensed for particular assays. Assays are started by adding assay buffer and reagents to the ARPs. This concept offers important logistical advantages for screening such as decoupling of the plate preparation from the screening process and exchange of assay plates between different geographical locations. Compound solutions can be accurately and precisely dispensed by acoustic droplet ejection technology in the small volumes required for screening in the 1536-well format. At such low volumes, however, potential effects such as solvent evaporation, compound degradation, precipitation, or adsorption are reasons for concern with regard to assay reproducibility, performance, and shelf life of ARPs. To address these concerns, the authors screened freshly prepared ARPs using several types of assays. The results were compared to results obtained from plates stored for up to 13 days under 2 storage conditions (22°C,-18°C). Tight correlations between results were found that indi_cated that temperature and time had very little influence on the assay performance for up to about 1 week storage time of the plates. In addition, using a time series of microphotographs of DMSO droplets, the authors visually confirmed that the sizes of the droplets in ARPs apparently do not change over 13 days under certain storage conditions.
机译:可用于测定的化合物板(ARPs)是密封的测定板,其中包含针对特定测定预先分配的筛选化合物的DMSO溶液。通过向ARP中添加测定缓冲液和试剂来开始测定。该概念为筛选提供了重要的后勤优势,例如将板制备物从筛选过程中分离出来,以及在不同地理位置之间交换化验板。通过声学液滴喷射技术,可以将1536孔格式的筛分所需的小体积化合物准确,精确地分配到溶液中。但是,在如此低的体积下,诸如溶剂蒸发,化合物降解,沉淀或吸附之类的潜在影响是关于ARPs的测定重现性,性能和保存期限的原因。为了解决这些问题,作者使用几种类型的检测方法筛选了新鲜制备的ARP。将结果与在2种存储条件(22°C,-18°C)下最多存储13天的平板的结果进行比较。结果之间的紧密相关性表明,温度和时间对板的最长约1周的保存时间对测定性能的影响很小。此外,使用DMSO液滴的时间序列显微照片,作者直观地确认了在某些存储条件下ARP中液滴的大小显然在13天之内没有变化。

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