ZnO-poly(methyl methacrylate) nanobeads for enriching and desalting low-abundant proteins followed by directly MALDI-TOF MS analysis
ZnO-poly(methyl methacrylate) nanobeads for enriching and desalting low-abundant proteins followed by directly MALDI-TOF MS analysis
复制标题
ZnO-聚（甲基丙烯酸甲酯）纳米珠用于富集和脱盐低丰度蛋白质，然后直接进行 MALDI-TOF MS 分析
DOI：
10.1021/ac801001b
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发表时间：
2008-09-01
影响因子：
7.4
通讯作者：
Yang, Pengyuan
中科院分区：
文献类型：
--
作者：
Shen, Wenwen;Xiong, Huanming;Yang, Pengyuan
建立了一种以核-壳结构氧化锌-聚甲基丙烯酸甲酯（ZnO-PMMA）纳米球为吸附剂的快速固相微萃取方法。这种快速、高富集效率和耐盐能力的方法依赖于核壳纳米球的结构。首先，PMMA壳的大表面积使得分散的纳米珠快速捕获样品，凭借ZnO-PMMA与样品之间的多种相互作用，除了与盐的相互作用。其次，ZnO核的小纳米尺寸（2.1 nm）和柔性疏水PMMA壳，可以防止核聚集，使纳米珠在基质辅助激光解吸/电离（MALDI）板上形成均匀的层，不妨碍基质和样品的共结晶。第三，ZnO芯还防止PMMA在质谱仪中碎裂和电离。在这篇文章中，类似于80%的牛血清白蛋白酶被富集ZnO-PMMA从100 μ L的溶液中在10分钟内孵育，固相可以直接通过MALDI质谱分析。质量强度可以增加5-10倍（ZnO-PMMA富集相对于冻干）。即使存在饱和NaCl（6.2 M）、饱和NH 4 HCO 3（2.6 M）或1 M尿素，也可以获得高质量的质谱。该方法已成功应用于人大肠癌蛋白质组研究，发现了8个新蛋白质。
A fast solid-phase microextraction method using core-shell ZnO-poly (methyl methacrylate) nanobeads (ZnO-PMMA) as adsorbent was established. This fast method with high enriching efficiency and salt tolerance capability depends on the structure of the core-shell nanobeads. First, the large surface area of the PMMA shell makes the dispersive nanobeads capture samples quickly, by virtue of multi-interactions between ZnO-PMMA and samples except for the interaction with salts. Second, the small nanosize of the ZnO-core (2.1 nm) and the flexible hydrophobic PMMA shell, which can prevent the cores from aggregation, make the nanobeads form a homogeneous layer on the matrix-assisted laser desorption/ionization (MALDI) plate and do not hinder the cocrystallization of the matrix and samples. Third, the ZnO core also prevents PMMA from fragmentation and ionization in mass spectrometer. In this article, similar to 80% bovine serum albumin digests were enriched by ZnO-PMMA from 100 amol/mu L solution within 10-min incubation, and the solid phase can be directly analyzed by MALDI mass spectrometry. Mass intensity can be increased 5-10-fold (ZnO-PMMA enrichment vs lyophilization). High-quality mass spectra can be obtained, even with the presence of saturated NaCl (6.2 M), saturated NH4HCO3 (2.6 M), or 1 M urea. This method has been successfully applied to human colorectal cancer proteome research, and eight new proteins have been found.