Center for Molecular Imaging, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, United States.
Anal Chem. 2013 Apr 2;85(7):3508-14. doi: 10.1021/ac303199x. Epub 2013 Mar 18.
The discovery of small molecule ligands targeted to the surface of live pathogenic bacteria would enable an entirely new class of antibiotics. We report the development and validation of a microarray-based high-throughput screening platform for bacteria that exploits 300 μm diameter chemical spots in a 1 in. × 3 in. nanolayered glass slide format. Using 24 model compounds and 4 different bacterial strains, we optimized the screening technology, including fluorophore-based optical deconvolution for automated scoring of affinity and cyan-magenta-yellow-key (CMYK) color-coding for scoring of both affinity and specificity. The latter provides a lossless, one-dimensional view of multidimensional data. By linking in silico analysis with cell binding affinity and specificity, we could also begin to identify the physicochemical factors that affect ligand performance. The technology we describe could form the foundation for developing new classes of antibiotics.
针对活病原细菌表面的小分子配体的发现将能够产生全新类别的抗生素。我们报告了一种基于微阵列的高通量筛选平台的开发和验证，该平台利用 1 英寸×3 英寸纳米层玻璃载玻片格式中的 300μm 直径化学斑点来筛选细菌。使用 24 种模型化合物和 4 种不同的细菌菌株，我们优化了筛选技术，包括基于荧光染料的光学离卷，用于自动评分亲和力，以及青色-品红色-黄色-关键色（CMYK）编码，用于评分亲和力和特异性。后者提供了多维数据的无损一维视图。通过将计算机分析与细胞结合亲和力和特异性相结合，我们还可以开始确定影响配体性能的物理化学因素。我们描述的技术可以为开发新类别的抗生素奠定基础。
