Poulsen Sally-Ann
Eskitis Institute for Cell and Molecular Therapies, Griffith University, Nathan, Brisbane, 4111, Queensland, Australia.
J Am Soc Mass Spectrom. 2006 Aug;17(8):1074-1080. doi: 10.1016/j.jasms.2006.03.017. Epub 2006 Jun 2.
A dynamic combinatorial library (DCL) screening approach is described that permits direct identification of the effective (from ineffective) combination of building blocks in the equilibrating DCL. The approach uses Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS) together with sustained off-resonance irradiation collision activated dissociation (SORI-CAD) to detect noncovalent protein-DCL ligand complexes under native conditions. It was shown that in a single, rapid experiment one could concurrently identify all the ligands of interest from the DCL against a background of inactive DCL ligands while still in the presence of the target protein. This result has demonstrated that mass spectrometry may provide a fast preliminary screening approach to identify DCL candidates for later verification with more traditional but time-consuming analysis. The MS/MS enables DCL mixtures to be effectively deconvoluted without the need for either chromatography, synthesis of DCL sub-libraries, conversion of the DCL to a static library, or disruption of the protein-ligand complexes before analysis--all typically necessary for the current screening method for DCLs.
本文描述了一种动态组合库（DCL）筛选方法，该方法能够直接在平衡的DCL中识别有效（与无效相对）的构建块组合。该方法使用傅里叶变换离子回旋共振质谱（FTICR MS）和持续非共振辐照碰撞激活解离（SORI-CAD）来在天然条件下检测非共价蛋白质-DCL配体复合物。结果表明，在单个快速实验中，人们可以在仍存在靶蛋白的情况下，在无活性DCL配体的背景下同时从DCL中识别出所有感兴趣的配体。这一结果表明，质谱法可为识别DCL候选物提供一种快速的初步筛选方法，以便随后用更传统但耗时的分析方法进行验证。串联质谱（MS/MS）能够有效地对DCL混合物进行解卷积，而无需进行色谱分析、合成DCL子库、将DCL转化为静态库或在分析前破坏蛋白质-配体复合物，而这些通常是当前DCL筛选方法所必需的。
