Ozhalici-Unal Hayriye, Pow Crystal Lee, Marks Sarah A, Jesper Lawrence D, Silva Gloria L, Shank Nathaniel I, Jones Elizabeth W, Burnette James M, Berget Peter B, Armitage Bruce A
Department of Chemistry, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213, USA.
J Am Chem Soc. 2008 Sep 24;130(38):12620-1. doi: 10.1021/ja805042p. Epub 2008 Aug 30.
Combined magnetic and fluorescence cell sorting were used to select Fluorogen Activating Proteins (FAPs) from a yeast surface-displayed library for binding to the fluorogenic cyanine dye Dimethyl Indole Red (DIR). Several FAPs were selected that bind to the dye with low nanomolar Kd values and enhance fluorescence more than 100-fold. One of these FAPs also exhibits considerable promiscuity, binding with high affinity to several other fluorogenic cyanine dyes with emission wavelengths covering most of the visible and near-IR regions of the spectrum. This significantly expands the number and wavelength range of scFv-based fluoromodules.
结合磁性和荧光细胞分选技术，从酵母表面展示文库中筛选出能与荧光花菁染料二甲基吲哚红（DIR）结合的荧光激活蛋白（FAP）。筛选出了几种FAP，它们与该染料的结合解离常数（Kd）值低至纳摩尔级别，且能使荧光增强100倍以上。其中一种FAP还表现出相当的通用性，能与其他几种发射波长覆盖光谱大部分可见光和近红外区域的荧光花菁染料高亲和力结合。这显著扩大了基于单链抗体片段（scFv）的荧光调节模块的数量和波长范围。
