当前位置：
X-MOL 学术
›
ACS Comb. Sci.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Off-DNA DNA-Encoded Library Affinity Screening.
ACS Combinatorial Science Pub Date : 2019-12-12 , DOI: 10.1021/acscombsci.9b00153 Amber L Hackler , Forrest G FitzGerald , Vuong Q Dang , Alexander L Satz 1 , Brian M Paegel
ACS Combinatorial Science Pub Date : 2019-12-12 , DOI: 10.1021/acscombsci.9b00153 Amber L Hackler , Forrest G FitzGerald , Vuong Q Dang , Alexander L Satz 1 , Brian M Paegel
Affiliation
|
DNA-encoded library (DEL) technology is emerging as a key element of the small molecule discovery toolbox. Conventional DEL screens (i.e., on-DNA screening) interrogate large combinatorial libraries via affinity selection of DNA-tagged library members that are ligands of a purified and immobilized protein target. In these selections, the DNA tags can materially and undesirably influence target binding and, therefore, the experiment outcome. Here, we use a solid-phase DEL and droplet-based microfluidic screening to separate the DEL member from its DNA tag (i.e., off-DNA screening), for subsequent in-droplet laser-induced fluorescence polarization (FP) detection of target binding, obviating DNA tag interference. Using the receptor tyrosine kinase (RTK) discoidin domain receptor 1 (DDR1) as a proof-of-concept target in a droplet-scale competition-binding assay, we screened a 67 100-member solid-phase DEL of drug-like small molecules for competitive ligands of DDR1 and identified several known RTK inhibitor pharmacophores, including azaindole- and quinazolinone-containing monomers. Off-DNA DEL affinity screening with FP detection is potentially amenable to a wide array of target classes, including nucleic acid binding proteins, proteins that are difficult to overexpress and purify, or targets with no known activity assay.
中文翻译：
非DNA DNA编码文库亲和力筛选。
DNA编码文库（DEL）技术正在成为小分子发现工具箱的关键要素。常规的DEL筛选（即DNA上筛选）通过亲和选择DNA标记的文库成员来查询大型组合文库，这些成员是纯化并固定的蛋白质靶标的配体。在这些选择中，DNA标签会实质性和不希望地影响靶标结合，从而影响实验结果。在这里，我们使用固相DEL和基于液滴的微流体筛选将DEL成员与其DNA标签分离（即脱DNA筛选），用于随后的液滴内激光诱导的荧光偏振（FP）检测靶标结合，避免了DNA标签的干扰。使用受体酪氨酸激酶（RTK）盘状蛋白结构域受体1（DDR1）作为液滴规模竞争结合测定中的概念证明目标，我们筛选了67 100个成员的药物样小分子固相DEL用于DDR1的竞争性配体，并鉴定了几种已知的RTK抑制剂药效基团，包括含氮吲哚和喹唑啉酮的单体。带有FP检测的DNA外DEL亲和力筛选可能适用于各种各样的目标类别，包括核酸结合蛋白，难以过表达和纯化的蛋白质或没有已知活性测定的目标。
更新日期：2019-12-31
中文翻译：
非DNA DNA编码文库亲和力筛选。
DNA编码文库（DEL）技术正在成为小分子发现工具箱的关键要素。常规的DEL筛选（即DNA上筛选）通过亲和选择DNA标记的文库成员来查询大型组合文库，这些成员是纯化并固定的蛋白质靶标的配体。在这些选择中，DNA标签会实质性和不希望地影响靶标结合，从而影响实验结果。在这里，我们使用固相DEL和基于液滴的微流体筛选将DEL成员与其DNA标签分离（即脱DNA筛选），用于随后的液滴内激光诱导的荧光偏振（FP）检测靶标结合，避免了DNA标签的干扰。使用受体酪氨酸激酶（RTK）盘状蛋白结构域受体1（DDR1）作为液滴规模竞争结合测定中的概念证明目标，我们筛选了67 100个成员的药物样小分子固相DEL用于DDR1的竞争性配体，并鉴定了几种已知的RTK抑制剂药效基团，包括含氮吲哚和喹唑啉酮的单体。带有FP检测的DNA外DEL亲和力筛选可能适用于各种各样的目标类别，包括核酸结合蛋白，难以过表达和纯化的蛋白质或没有已知活性测定的目标。
京公网安备 11010802027423号