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基于数据非依赖采集的高通量文库生成和生物标志物检测的自动化蛋白质组学工作流程。

Automated Proteomics Workflows for High-Throughput Library Generation and Biomarker Detection Using Data-Independent Acquisition.

机构信息

School of Veterinary Science, The University of Queensland, Gatton, QLD 4343, Australia.

Central Analytical Research Facility, Queensland University of Technology, Brisbane, QLD 4001, Australia.

出版信息

J Proteome Res. 2023 Jun 2;22(6):2018-2029. doi: 10.1021/acs.jproteome.3c00074. Epub 2023 May 23.

Abstract

Sequential window acquisition of all theoretical mass spectra-mass spectrometry underpinned by advanced bioinformatics offers a framework for comprehensive analysis of proteomes and the discovery of robust biomarkers. However, the lack of a generic sample preparation platform to tackle the heterogeneity of material collected from different sources may be a limiting factor to the broad application of this technique. We have developed universal and fully automated workflows using a robotic sample preparation platform, which enabled in-depth and reproducible proteome coverage and characterization of bovine and ovine specimens representing healthy animals and a model of myocardial infarction. High correlation ( = 0.85) between sheep proteomics and transcriptomics datasets validated the developments. The findings suggest that automated workflows can be employed for various clinical applications across different animal species and animal models of health and disease.

摘要

序贯窗口采集所有理论质谱-先进的生物信息学支持的质谱分析为全面分析蛋白质组学和发现稳健的生物标志物提供了一个框架。然而,缺乏通用的样品制备平台来解决从不同来源收集的材料的异质性,可能是限制该技术广泛应用的一个因素。我们使用机器人样品制备平台开发了通用的、全自动的工作流程,该平台能够深入、可重复地覆盖和表征来自健康动物和心肌梗死模型的牛和羊标本的蛋白质组。绵羊蛋白质组学和转录组学数据集之间的高相关性(= 0.85)验证了这些发展。研究结果表明,自动化工作流程可用于不同动物物种和健康与疾病动物模型的各种临床应用。

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