دورية أكاديمية
Label-free LC-MS based assay to characterize small molecule compound binding to cells
| العنوان: | Label-free LC-MS based assay to characterize small molecule compound binding to cells |
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| المؤلفون: | Menglin Chen, Zhengrong Zhu, Tom Wisniewski, Xiaoping Zhang, David G. McLaren, Adam Weinglass, S. Adrian Saldanha |
| المصدر: | SLAS Discovery, Vol 27, Iss 7, Pp 405-412 (2022) |
| بيانات النشر: | Elsevier, 2022. |
| سنة النشر: | 2022 |
| المجموعة: | LCC:Medicine (General) LCC:Biotechnology |
| مصطلحات موضوعية: | Label-free technologies, LC-MS, Cell-based assays, Affinity binding, Medicine (General), R5-920, Biotechnology, TP248.13-248.65 |
| الوصف: | Study of small molecule binding to live cells provides important information on the characterization of ligands pharmacologically. Here we developed and validated a label-free, liquid chromatography-mass spectrometry (LC-MS) based cell binding assay, using centrifugation to separate binders from non-binders. This assay was applied to various target classes, with particular emphasis on those for which protein-based binding assay can be difficult to achieve. In one example, to study a G protein coupled receptor (GPCR), we used one antagonist as probe and multiple other antagonists as competitor ligands. Binding of the probe was confirmed to be specific and saturable, reaching a fast equilibrium. Competition binding analysis by titration of five known ligands suggested a good correlation with their inhibition potency. In another example, this assay was applied to an ion channel target with its agonists, of which the determined binding affinity was consistent with functional assays. This versatile method allows quantitative characterization of ligand binding to cell surface expressed targets in a physiologically relevant environment. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2472-5552 |
| Relation: | http://www.sciencedirect.com/science/article/pii/S2472555222136920; https://doaj.org/toc/2472-5552 |
| DOI: | 10.1016/j.slasd.2022.08.005 |
| URL الوصول: | https://doaj.org/article/692086835e5a4837a6ecf8d1a559ee73 |
| رقم الأكسشن: | edsdoj.692086835e5a4837a6ecf8d1a559ee73 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 24725552 |
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| DOI: | 10.1016/j.slasd.2022.08.005 |