One-Pot Time-Induced Proteome Integral Solubility Alteration Assay for Automated and Sensitive Drug–Target Identification

IF 6.7 1区 化学 Q1 CHEMISTRY, ANALYTICAL Analytical Chemistry Pub Date : 2024-11-20 DOI:10.1021/acs.analchem.4c05127
Zhaowei Meng, Amir Ata Saei, Hezheng Lyu, Massimiliano Gaetani, Roman A. Zubarev
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Abstract

The proteome integral solubility alteration (PISA) assay is widely used for identifying drug targets, but it is labor-intensive and time-consuming and requires a substantial amount of biological sample. Aiming at enabling automation and greatly reducing the sample amount, we developed one-pot time-induced (OPTI)-PISA. Here, we demonstrate OPTI-PISA performance on identifying targets of multiple drugs in cell lysate and scaling down the sample amount to sub-microgram levels, making the PISA method suitable for NanoProteomics. OPTI-PISA can be implemented using only the standard equipment of a proteomics lab.

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用于自动灵敏鉴定药物靶点的单锅时间诱导蛋白质组整体溶解度改变分析法
蛋白质组整体溶解度改变(PISA)测定法被广泛用于鉴定药物靶点,但该方法耗时耗力,需要大量生物样本。为了实现自动化并大大减少样本量,我们开发了单锅时间诱导(OPTI)-PISA。在这里,我们展示了 OPTI-PISA 在鉴定细胞裂解物中多种药物靶点方面的性能,并将样品量减少到亚微克级,使 PISA 方法适用于纳米蛋白质组学。OPTI-PISA 只需使用蛋白质组学实验室的标准设备即可实现。
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来源期刊
Analytical Chemistry
Analytical Chemistry 化学-分析化学
CiteScore
12.10
自引率
12.20%
发文量
1949
审稿时长
1.4 months
期刊介绍: Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.
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