Kolářová Lenka, Kučera Lukáš, Vaňhara Petr, Hampl Aleš, Havel Josef
Department of Chemistry, Faculty of Science, Masaryk University, Kamenice 5/A14, 62500, Brno, Czech Republic.
Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Kamenice 3/A1, 62500, Brno, Czech Republic.
Rapid Commun Mass Spectrom. 2015 Sep 15;29(17):1585-1595. doi: 10.1002/rcm.7265.
Many kinds of nanoparticles (NPs) have been used for mass spectrometry (MS) so far. Here we report the first use of flower-like gold nanoparticles (AuNPs) as a mediator to enhance ionization in MS of peptides and proteins.
Flower-like AuNPs were characterized using transmission and scanning electron microscopy, UV-VIS spectrophotometry, and laser desorption/ionization (LDI)-MS and compared with polyhedral AuNPs. Mass spectra were obtained in positive ion mode using a time-of-flight (TOF) analyzer coupled with either matrix-assisted laser desorption/ionization (MALDI) or surface-assisted laser desorption/ionization (SALDI) methods.
The intensities of peptide peaks (m/z 500-3500) were up to 7.5× and up to 7× higher using flower-like AuNPs and flower-like AuNPs-enriched α-cyano-4-hydroxycinnamic acid (CHCA) matrix respectively, than the classical CHCA matrix. The signals of higher mass peptide/protein peaks (m/z 3600-17000) were up to 2× higher with using flower-like AuNPs-enriched CHCA matrix than conventional CHCA matrix. The signal of profile peaks generated by intact cell MALDI-TOFMS of fibroblast suspension (m/z 4000-20000) was 2× higher with using flower-like AuNPs combined with sinapinic acid (SA) compared to SA matrix alone. The use of flower-like AuNPs as internal calibration standard for the calibration of MS spectra of peptides was performed.
Flower-like AuNPs and flower-like AuNPs combined with CHCA or SA as combined matrices for MS measurement of peptides and proteins were used. Comparison of the conventional MALDI method and our method with flower-like AuNPs was carried out. In addition, gold clusters generated from flower-like AuNPs by SALDI provide a suitable internal calibration standard for MS analysis of peptides. Copyright © 2015 John Wiley & Sons, Ltd.
到目前为止,多种纳米颗粒(NPs)已被用于质谱分析(MS)。在此,我们首次报道使用花状金纳米颗粒(AuNPs)作为介质来增强肽和蛋白质质谱分析中的离子化。
使用透射和扫描电子显微镜、紫外可见分光光度法以及激光解吸/电离(LDI)-MS对花状AuNPs进行表征,并与多面体AuNPs进行比较。使用飞行时间(TOF)分析仪在正离子模式下通过基质辅助激光解吸/电离(MALDI)或表面辅助激光解吸/电离(SALDI)方法获得质谱图。
使用花状AuNPs和富含花状AuNPs的α-氰基-4-羟基肉桂酸(CHCA)基质时,肽峰(m/z 500 - 3500)的强度分别比经典CHCA基质高出7.5倍和7倍。使用富含花状AuNPs的CHCA基质时,较高质量肽/蛋白质峰(m/z 3600 - 17000)的信号比传统CHCA基质高出2倍。与单独使用芥子酸(SA)基质相比,使用花状AuNPs与芥子酸(SA)结合时,成纤维细胞悬浮液的完整细胞MALDI-TOFMS产生的轮廓峰信号(m/z 4000 - 20000)高出2倍。还进行了使用花状AuNPs作为肽质谱图校准的内标。
使用花状AuNPs以及花状AuNPs与CHCA或SA结合作为肽和蛋白质质谱测量的组合基质。对传统MALDI方法和我们使用花状AuNPs方法进行了比较。此外,通过SALDI从花状AuNPs产生的金簇为肽的质谱分析提供了合适的内标。版权所有© 2015约翰威立父子有限公司。