Mass spectrometry signal amplification for ultrasensitive glycoprotein detection using gold nanoparticle as mass tag combined with boronic acid based isolation strategy.
Mass spectrometry signal amplification for ultrasensitive glycoprotein detection using gold nanoparticle as mass tag combined with boronic acid based isolation strategy.
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DOI：
10.1016/j.aca.2013.05.063
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发表时间：
2013-07
影响因子：
6.2
通讯作者：
Minbo Liu;Lijuan Zhang;Yawei Xu;Pengyuan Yang;Haojie Lu
中科院分区：
文献类型：
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作者：
Minbo Liu;Lijuan Zhang;Yawei Xu;Pengyuan Yang;Haojie Lu
我们描述了一种新的方法，用于快速和超灵敏检测完整的糖蛋白，而无需酶预处理，这是常用的蛋白质组学研究。该方法以金纳米粒子（AuNP）为信号标记物，结合硼酸辅助分离策略，实现了激光解吸/电离质谱（LDI-MS）分析。简单地说，首先用硼酸功能化的磁性微粒从样品溶液中分离目标糖蛋白，然后加入表面修饰的金纳米颗粒以共价结合糖蛋白。之后，将这些AuNP标记的糖蛋白从磁性微粒洗脱并应用于LDI-MS分析。该策略检测并记录的是金纳米粒子的质量信号，而不是糖蛋白的质量信号。通过对不同标准糖蛋白的数据处理，我们证明了金纳米粒子的信号可以用来定量表征糖蛋白。该方法允许飞摩尔检测完整的糖蛋白。我们相信，这种方法的成功验证三种不同类型的糖蛋白表明，在不久的将来用于跟踪痕量的目标糖蛋白在真实的生物样品中的潜在用途。
We describe a novel method for rapid and ultrasensitive detection of intact glycoproteins without enzymatic pretreatment which was commonly used in proteomic research. This method is based on using gold nanoparticle (AuNP) as signal tag in laser desorption/ionization mass spectrometry (LDI-MS) analysis combined with boronic acid assisted isolation strategy. Briefly speaking, target glycoproteins were firstly isolated from sample solution with boronic acid functionalized magnetic microparticles, and then the surface modified gold nanoparticles were added to covalently bind to the glycoproteins. After that, these AuNP tagged glycoproteins were eluted from magnetic microparticles and applied to LDI-MS analysis. The mass signal of AuNP rather than that of glycoprotein was detected and recorded in this strategy. Through data processing of different standard glycoproteins, we have demonstrated that the signal of AuNP could be used to quantitatively represent glycoprotein. This method allows femtomolar detection of intact glycoproteins. We believe that the successful validation of this method on three different kinds of glycoproteins suggests the potential use for tracking trace amount of target glycoproteins in real biological samples in the near future.