Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan.
Anal Chem. 2010 Jul 15;82(14):5951-7. doi: 10.1021/ac101426n.
Although matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) analysis is an important tool for analyzing and characterizing biomolecules of varying complexity, the sensitivity of MALDI-TOFMS is dependent on proper preparation of the sample, a process that is oftentimes problematic and requires considerable expertise. In this study, we have developed a radiate microstructure chip on which samples can be concentrated for analysis by MALDI-TOFMS. The sample/matrix mixture was deposited onto the central space of the well on the chip and allowed to dry. Microscopic analysis confirmed that the applied samples were confined to the central zone. Sample spots focused on the chip were much smaller than those on an unmodified plate with the same total volume. Optimizing processes of several preparation factors were also performed to ensure matrix homogeneity in our chip. Analysis of the samples with MALDI-TOFMS showed that the signals from samples on our chip were significantly greater than those on the unmodified plate. The feasibility of using this chip to detect peptides and phosphopeptides was also demonstrated.
尽管基质辅助激光解吸/电离飞行时间质谱(MALDI-TOFMS)分析是分析和鉴定具有不同复杂性的生物分子的重要工具,但 MALDI-TOFMS 的灵敏度取决于样品的正确制备,这一过程常常存在问题,需要相当的专业知识。在这项研究中,我们开发了一种放射状微结构芯片,可将样品浓缩进行 MALDI-TOFMS 分析。将样品/基质混合物沉积在芯片上的孔中央空间并使其干燥。显微镜分析证实,施加的样品被限制在中央区域。聚焦在芯片上的样品点明显小于具有相同总体积的未修饰板上的样品点。还对几个制备因素的优化过程进行了优化,以确保我们的芯片中基质的均匀性。使用 MALDI-TOFMS 对样品进行分析表明,来自我们芯片上的样品的信号明显大于来自未修饰板上的样品的信号。还证明了使用该芯片检测肽和磷酸肽的可行性。