Selection of bead-displayed, PNA-encoded chemicals.
Selection of bead-displayed, PNA-encoded chemicals.
复制标题
选择珠子展示、PNA 编码的化学品。
DOI：
10.1002/jmr.1007
复制
发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Macosko,JedC
中科院分区：
文献类型：
--
作者：
Gassman,NatalieR;Nelli,JPatrick;Dutta,Samrat;Kuhn,Adam;Bonin,Keith;Pianowski,Zbigniew;Winssinger,Nicolas;Guthold,Martin;Macosko,JedC
特定分子结合剂缺乏有效的识别和分离方法从根本上限制了药物的发现。在这里，我们开发了一种从组合生成的文库中选择具有特定功能特性的肽核酸（PNA）编码分子的方法。该方法由三个基本阶段组成：(1) 创建 Lab-on-Bead™ 文库，即单珠单序列文库，依次显示候选分子文库，(2) 荧光显微镜辅助识别单个靶标结合珠子，并通过微量移液器操纵器提取（湿法或干法）这些珠子及其附着的候选分子，以及 (3) 通过扩增和测序鉴定靶标结合候选分子。这种新颖的技术整合利用 DNA 检测方法的敏感性以及分子筛选的多重化和小型化性质，从大型核酸编码的化学库中有效地选择和识别靶标结合分子。除了加速目前用于发现新候选药物的测定的潜力之外，其简单的基于珠子的设计还可以轻松筛选各种准备好的表面，从而可以将该技术的应用扩展到诊断试剂和疾病标记物的发现。版权所有 © 2009 约翰·威利父子有限公司
The lack of efficient identification and isolation methods for specific molecular binders has fundamentally limited drug discovery. Here, we have developed a method to select peptide nucleic acid (PNA) encoded molecules with specific functional properties from combinatorially generated libraries. This method consists of three essential stages: (1) creation of a Lab‐on‐Bead™ library, a one‐bead, one‐sequence library that, in turn, displays a library of candidate molecules, (2) fluorescence microscopy‐aided identification of single target‐bound beads and the extraction – wet or dry – of these beads and their attached candidate molecules by a micropipette manipulator, and (3) identification of the target‐binding candidate molecules via amplification and sequencing. This novel integration of techniques harnesses the sensitivity of DNA detection methods and the multiplexed and miniaturized nature of molecule screening to efficiently select and identify target‐binding molecules from large nucleic acid encoded chemical libraries. Beyond its potential to accelerate assays currently used for the discovery of new drug candidates, its simple bead‐based design allows for easy screening over a variety of prepared surfaces that can extend this technique's application to the discovery of diagnostic reagents and disease markers. Copyright © 2009 John Wiley & Sons, Ltd.