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用于低输入蛋白质组学中样品制备的数字微流控技术。

Digital Microfluidics for Sample Preparation in Low-Input Proteomics.

作者信息

Steinbach Max K, Leipert Jan, Matzanke Theo, Tholey Andreas

机构信息

Systematic Proteome Research & Bioanalytics, Institute for Experimental Medicine, Christian-Albrechts-Universität zu Kiel, 24105, Kiel, Germany.

出版信息

Small Methods. 2025 Jan;9(1):e2400495. doi: 10.1002/smtd.202400495. Epub 2024 Aug 29.

Abstract

Low-input proteomics, also referred to as micro- or nanoproteomics, has become increasingly popular as it allows one to elucidate molecular processes in rare biological materials. A major prerequisite for the analytics of minute protein amounts, e.g., derived from low cell numbers, down to single cells, is the availability of efficient sample preparation methods. Digital microfluidics (DMF), a technology allowing the handling and manipulation of low liquid volumes, has recently been shown to be a powerful and versatile tool to address the challenges in low-input proteomics. Here, an overview is provided on recent advances in proteomics sample preparation using DMF. In particular, the capability of DMF to isolate proteomes from cells and small model organisms, and to perform all necessary chemical sample preparation steps, such as protein denaturation and proteolytic digestion on-chip, are highlighted. Additionally, major prerequisites to making these steps compatible with follow-up analytical methods such as liquid chromatography-mass spectrometry will be discussed.

摘要

低输入蛋白质组学,也被称为微量或纳升蛋白质组学,因其能够阐明稀有生物材料中的分子过程而越来越受欢迎。对微量蛋白质(例如源自少量细胞甚至单细胞)进行分析的一个主要前提是要有高效的样品制备方法。数字微流控(DMF)是一种能够处理和操控低液体体积的技术,最近已被证明是应对低输入蛋白质组学挑战的强大且通用的工具。本文概述了使用DMF进行蛋白质组学样品制备的最新进展。特别强调了DMF从细胞和小型模式生物中分离蛋白质组以及在芯片上执行所有必要化学样品制备步骤(如蛋白质变性和蛋白水解消化)的能力。此外,还将讨论使这些步骤与后续分析方法(如液相色谱 - 质谱联用)兼容的主要前提条件。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9d4/11740955/cef7a70ce184/SMTD-9-2400495-g013.jpg

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