Mayr L M
Novartis Institutes of BioMedical Research, Discovery Technologies/Lead Discovery Center, Basel, Switzerland.
Ernst Schering Res Found Workshop. 2006(58):111-73. doi: 10.1007/978-3-540-37635-4_8.
Drug discovery in the chemogenomic space has seen some tremendous changes over the last decade. Compared to previous times, not only the number of available chemical compounds for screening, but also the number of molecular targets used for screening has increased significantly. This has triggered the need for very fast, efficient, and effective novel readout technologies for compound testing. Novartis has developed two novel high-throughput screening (HTS) technologies for that purpose--NanoScreen and SpeedScreen. NanoScreen is a highly miniaturized and fully automated HTS/uHTS test system with confocal single-molecule as well as non-confocal detection capabilities and is used for functional screening in the range of 1-5 microl per sample. The integration of the single-molecule readout technologies into the system enables highly sophisticated biochemical test systems with multi-parameter readout for very high data quality. SpeedScreen is a highly miniaturized and automated screening system for high-throughput affinity-selection of compounds. In practice, pools of compounds are incubated with the target protein and the unbound chemical compounds are removed from the target-compound complex via very fast, multiparallel size-exclusion-chromatography. The holoenzyme is disintegrated and analyzed via microbore reversed-phase high performance liquid chromatography (microbore RP-HPLC). Both systems have been developed and implemented with great success at the Novartis Lead Discovery Center (LDC) in Basel. These technologies have enabled us to access targets that would otherwise not have been possible, e.g., very expensive targets, "orphan" drug targets, or targets that are "non-tractable" by conventional screening technologies. Taken together, these novel screening technologies enable novel approaches for chemogenomic research that would have not been possible in the past.
在过去十年中，化学基因组学领域的药物发现发生了一些巨大的变化。与以往相比，不仅用于筛选的可用化合物数量增加了，而且用于筛选的分子靶点数量也显著增加。这引发了对用于化合物测试的非常快速、高效且有效的新型读出技术的需求。诺华公司为此开发了两种新型高通量筛选（HTS）技术——纳米筛选（NanoScreen）和快速筛选（SpeedScreen）。纳米筛选是一种高度微型化且完全自动化的HTS/超高通量筛选（uHTS）测试系统，具有共聚焦单分子以及非共聚焦检测能力，用于每个样品1 - 5微升范围内的功能筛选。将单分子读出技术集成到该系统中，可实现具有多参数读出的高度复杂的生化测试系统，以获得非常高的数据质量。快速筛选是一种高度微型化且自动化的筛选系统，用于化合物的高通量亲和选择。在实际操作中，将化合物库与目标蛋白一起孵育，然后通过非常快速的多平行尺寸排阻色谱法从目标 - 化合物复合物中去除未结合的化合物。全酶经微径反相高效液相色谱（微径RP - HPLC）分解并分析。这两种系统均已在巴塞尔的诺华公司先导化合物发现中心（LDC）成功开发并实施。这些技术使我们能够接触到那些否则不可能接触到的靶点，例如非常昂贵的靶点、“孤儿”药物靶点或传统筛选技术“难以处理”的靶点。综上所述，这些新型筛选技术为化学基因组学研究带来了过去不可能实现的新方法。
