自动化高通量亲和度捕获-质谱平台与数据独立获取
Hui Jing1, Paul L Richardson1, Gregory K Potts1
1Discovery Research, AbbVie, Inc., 1 North Waukegan Rd., North Chicago, Illinois 60064, United States.
Journal of proteome research
|January 27, 2025
概括
我们开发了一种自动化的亲和度捕获-质谱工作流程,用于高通量药物标分析. 这种方法显著减少了样本准备时间,并提高了小分子选择性研究的激酶覆盖率.
科学领域:
- 化学蛋白质组学 化学蛋白质组学
- 药物发现 药物发现
- 质谱测量质量谱测量
背景情况:
- 与质谱学 (MS) 结合的亲和度捕获 (AC) 对于评估药物发现中小分子标参与度和选择性至关重要.
- 当前的AC-MS方法在高通量应用中面临局限性,原因是样本准备和MS获取的时间很长.
- 需要简化,自动化工作流程来加速目标参与研究.
研究的目的:
- 开发和验证一个自动化,高通量AC-MS工作流程,用于小分子标造型.
- 实施数据独立获取 (DIA) -MS方法,以增强基因组覆盖和定量分析.
- 为了证明工作流的实用性,在评估药物选择性对广泛的激酶.
主要方法:
- 一个自动化的96井板交流工作流程,使用生物化探头和斯特雷普塔维丁磁珠进行目标丰富.
- 从EvoSep固体相提取方法直接采样,用于液体染色学-并联质谱 (LC-MS/MS) 分析.
- 开发一种无标签的定量DIA-MS方法,用于基因组分析.
主要成果:
- 自动化工作流显著减少了整体和实践样品准备时间.
- DIA-MS实现了大约380个激酶的覆盖,比数据依赖获取 (DDA-MS) 增加了60%以上.
- 证明了达沙替尼的可重复性标概况以及对两种CDK9抑制剂对约250种激酶的选择性评估.
结论:
- 开发的自动化AC-MS工作流程可以实现高效和高通量目标参与和选择性分析.
- 与传统方法相比,这种方法显著改善了激酶覆盖范围,并减少了分析时间.
- 工作流程为加速药物发现和开发管道提供了有价值的路线图.
更多相关视频
相关概念视频
Tandem Mass Spectrometry
878
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and signal-to-noise ratio for the analyte. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.
Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called collision-induced...
Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called collision-induced...
878
Peptide Identification Using Tandem Mass Spectrometry
6.4K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
6.4K
MALDI-TOF Mass Spectrometry
4.7K
Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.
Matrix-assisted laser desorption ionization (MALDI) is a commonly...
Matrix-assisted laser desorption ionization (MALDI) is a commonly...
4.7K
Mass Spectrometry: Complex Analysis
699
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
699
Mass Spectrometers
5.1K
This lesson details the instrumentation of a mass spectrometer—a physical instrument to perform mass spectrometry on analyte molecules and record the characteristic mass spectra. This is achieved via three chief functions:
5.1K
Mass Analyzers: Overview
569
The mass analyzer is a crucial component of the mass spectrometer. In the ionization chamber, the vaporized sample is bombarded with a high-energy electron beam to generate a radical cation and further fragment into neutral molecules, radicals, and cations. A series of negatively charged accelerator plates accelerate the cations into the mass analyzer. The mass analyzer separates ions according to their mass-to-charge (m/z) ratios and then directs them to the detector. The common types of mass...
569


