Ultra-High-Throughput and Low-Volume Analysis of Intact Proteins with LAP-MALDI MS.

Ultra-High-Throughput and Low-Volume Analysis of Intact Proteins with LAP-MALDI MS.
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LAP-MALDI MS用于完整蛋白质的超高容量和低容量分析。

DOI:
10.1021/jasms.3c00068
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发表时间:
2023-06-07
影响因子:
3.2
通讯作者:
Cramer, Rainer
Cramer, Rainer
中科院分区:
化学3区
文献类型:
--
作者:
Challen, Bob;Morris, Michael;Cramer, Rainer

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高通量(HTP)质谱学(MS)是一个快速发展的领域,随着许多技术的发展,以适应不断增加的样品分析速度。其中许多技术,如AEMS和IR-Maldesi MS,需要至少20-50μL才能进行分析。在这里,液体大气压基质辅助激光解吸电离(LAP-MALDIMS)被提出作为一种替代方案,用于在0.5μL液滴中只需要飞摩尔量的蛋白质就可以超高通量地分析蛋白质。通过用高速XY工作台驱动器移动384孔微滴定样品板,在每次扫描200个光谱的数据采集速率下,实现了每秒高达10个样品的样品采集速率。结果表明,LAP-MALDIMS可以分析浓度为≤2μM的混合蛋白质溶液,也可以分析浓度为0.2≤M的单个蛋白质溶液。因此,LAP-MALDIMS为多重μ蛋白质分析提供了一个很有前途的平台。
High-throughput (HTP) mass spectrometry (MS) is a rapidly growing field, with many techniques evolving to accommodate ever increasing sample analysis rates. Many of these techniques, such as AEMS and IR-MALDESI MS, require volumes of at least 20–50 μL for analysis. Here, liquid atmospheric pressure-matrix-assisted laser desorption/ionization (LAP-MALDI) MS is presented as an alternative for ultra-high-throughput analysis of proteins requiring only femtomole quantities of protein in 0.5 μL droplets. By moving a 384-well microtiter sample plate with a high-speed XY-stage actuator, sample acquisition rates of up to 10 samples per second have been achieved at a data acquisition rate of 200 spectra per scan. It is shown that protein mixture solutions with concentrations of ≤2 μM can be analyzed at this speed, while individual protein solutions can be analyzed at concentrations of ≤0.2 μM. Thus, LAP-MALDI MS provides a promising platform for multiplexed HTP protein analysis.
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