COMBINES-CID: An Efficient Method for De Novo Engineering of Highly Specific Chemically Induced Protein Dimerization Systems
COMBINES-CID: An Efficient Method for De Novo Engineering of Highly Specific Chemically Induced Protein Dimerization Systems
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DOI：
10.1021/jacs.9b03522
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发表时间：
2019-07-17
影响因子：
15
通讯作者：
Gu, Liangcai
中科院分区：
文献类型：
--
作者：
Kang, Shoukai;Davidsen, Kristian;Gu, Liangcai
化学诱导二聚化（CID）系统，其中两种蛋白质仅在小分子配体存在下二聚化，为小分子传感和致动提供了通用工具。然而，只有少数CID系统存在，并且创建对任何给定配体具有所需灵敏度和特异性的CID系统是一个未解决的问题。在这里，我们开发了一种广泛适用于不同配体的组合结合剂使能的CID选择（COMBINES-CID）方法。我们通过产生由大麻二酚诱导的具有高配体选择性的基于纳米抗体的异源二聚化系统来证明原理。我们将CID系统应用于体液中大麻二酚的灵敏夹心酶联免疫吸附测定，检测限为0.25 ng/mL。COMBINES-CID为扩展小分子检测的生物传感器工具包提供了一种高效、经济的解决方案。
Chemically induced dimerization (CID) systems, in which two proteins dimerize only in the presence of a small molecule ligand, offer versatile tools for small molecule sensing and actuation. However, only a handful of CID systems exist and creating one with the desired sensitivity and specificity for any given ligand is an unsolved problem. Here, we developed a combinatorial binders-enabled selection of CID (COMBINES-CID) method broadly applicable to different ligands. We demonstrated a proof-of-principle by generating nanobody-based heterodimerization systems induced by cannabidiol with high ligand selectivity. We applied the CID system to a sensitive sandwich enzyme-linked immunosorbent assay-like assay of cannabidiol in body fluids with a detection limit of similar to 0.25 ng/mL. COMBINES-CID provides an efficient, cost-effective solution for expanding the biosensor toolkit for small molecule detection.