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用于蛋白质组学样本自动鉴定的硅微结构和薄膜基质辅助激光解吸电离靶板的开发。

Development of silicon microstructures and thin-film MALDI target plates for automated proteomics sample identifications.

作者信息

Miliotis T, Marko-Varga G, Nilsson J, Laurell T

机构信息

Department of Analytical Chemistry, Lund University, Box 124, SE-221 00, Lund, Sweden.

出版信息

J Neurosci Methods. 2001 Aug 15;109(1):41-6. doi: 10.1016/s0165-0270(01)00399-5.

Abstract

Here we report on the development of a proteomic platform utilizing a piezoelectric flow-through dispensing unit made from silicon microstructures. The use of a novel surface coating, where matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI MS) targets were uniformly precoated with a thin film of matrix/nitrocellulose, made the sample preparation straightforward and enabled the enrichment and analysis of proteins at low levels in proteomics samples. We demonstrate this by analyzing excised spots in a biological sample originating from a human fetal fibroblast cell line that was subjected to 2D gel-electrophoresis. Furthermore, a sample deposition rate below 30 Hz results in an increased analyte density on the dispensed sample spot, rendering signal amplification. In general, the sensitivity for proteins and peptides can be enhanced 10-50 times compared to traditional MALDI sample preparation techniques.

摘要

在此,我们报告一种蛋白质组学平台的开发,该平台利用由硅微结构制成的压电流通式分配装置。使用一种新型表面涂层,即基质辅助激光解吸/电离飞行时间质谱(MALDI MS)靶点均匀预涂有一层基质/硝酸纤维素薄膜,使得样品制备变得简单,并能够在蛋白质组学样品中对低水平蛋白质进行富集和分析。我们通过分析来自人胎儿成纤维细胞系的生物样品中经二维凝胶电泳分离出的斑点来证明这一点。此外,低于30 Hz的样品沉积速率会导致分配的样品斑点上分析物密度增加,从而实现信号放大。一般来说,与传统的MALDI样品制备技术相比,蛋白质和肽的灵敏度可提高10至50倍。

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