基于 Micro-SPE 的自下而上蛋白质组综合工作流程,可对有限样本进行灵敏分析。

Q4 Biochemistry, Genetics and Molecular Biology
Alan J Zimmerman, Michal Greguš, Alexander R Ivanov
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引用次数: 0

摘要

临床和生物样本通常稀少而珍贵(如稀有细胞分离、微针组织活检、小容量液体活检,甚至单细胞或细胞器)。典型的大规模蛋白质组学方法分析的蛋白质量要高得多,但由于其与pg-、ng-和低μg级蛋白质样本量不兼容,因此不能直接用于有限样本的分析。为解决这一难题,我们在此报告了基于微固相萃取头(OmSET)的样品制备工作流程,用于有限生物样品的灵敏分析。所开发的平台已成功用于 100-10,000 个典型哺乳动物细胞的分析测试,并可进行扩展,以便分析更低或更多的蛋白质量和更多的样品(即更高的分析通量)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comprehensive Micro-SPE-Based Bottom-Up Proteomic Workflow for Sensitive Analysis of Limited Samples.

Clinical and biological samples are often scarce and precious (e.g., rare cell isolates, microneedle tissue biopsies, small-volume liquid biopsies, and even single cells or organelles). Typical large-scale proteomic methods, where significantly higher protein amounts are analyzed, are not directly transferable to the analysis of limited samples due to their incompatibility with pg-, ng-, and low-μg-level protein sample amounts. Here, we report the on-microsolid-phase extraction tip (OmSET)-based sample preparation workflow for sensitive analysis of limited biological samples to address this challenge. The developed platform was successfully tested for the analysis of 100-10,000 typical mammalian cells and is scalable to allow for lower and larger protein amounts and more samples to be analyzed (i.e., higher throughput of analysis).

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来源期刊
Methods in molecular biology
Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
2.00
自引率
0.00%
发文量
3536
期刊介绍: For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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