Self-assembled monolayers for MALDI-TOF mass Spectrometry for Immunoassays of human protein antigens
Self-assembled monolayers for MALDI-TOF mass Spectrometry for Immunoassays of human protein antigens
复制标题
DOI：
10.1021/ac0701738
复制
发表时间：
2007-08-01
影响因子：
7.4
通讯作者：
Mrksich, Milan
中科院分区：
文献类型：
--
作者：
Patrie, Steven M.;Mrksich, Milan
本文报道了一种结合自组装单分子膜和基质辅助激光解吸/电离飞行时间质谱对临床样品进行免疫测定的方法。免疫传感器;通过将His标记的蛋白G（或A）固定到呈现Ni 2+螯合物的单层上，然后固定对预期分析物具有特异性的IgG抗体来制备。SAMDI质谱技术证实了免疫传感器上两种蛋白质的存在，并另外提供了结合到传感器的抗原的无标记分析。本文报道了检测人血清中几种蛋白质的实例，包括根据SAMDI光谱中的质荷比分辨每种分析物的多分析物测定。描述了一个实例，其中SAMDI用于鉴定来自诊断患有多发性硬化的患者的脑脊液中的胱抑素C的蛋白水解片段。SAMDI-TOF免疫测定结合了用于分析物的选择性和可再现定位的明确定义的表面化学与用于分析物的无标记检测的质谱法，可以提供替代方法来解决与标准化临床诊断相关的许多问题。
This paper reports a method that combines self-assembled monolayers; with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to perform immunoassays on clinical samples. The immunosensors; are prepared by immobilizing His-tagged protein G (or A) to a monolayer presenting the Ni2+ chelates, followed by immobilization of IgG antibodies with specificity for the intended analyte. The SAMDI mass spectrometry technique confirms the presence of the two proteins on the immunosensor and additionally provides a label-free analysis of antigens that bind to the sensor. This paper reports examples of detecting several proteins from human serum, including multianalyte assays that resolve each analyte according to their mass-to-charge ratio in the SAMDI spectra. An example is described wherein SAMDI is used to identify a proteolytic fragment of cystatin C in cerebral spinal fluids from patients diagnosed with multiple sclerosis. The SAMDI-TOF immunoassay, which combines well-defined surface chemistries for the selective and reproducible localization of analytes with mass spectrometry for label-free detection of analytes, may offer an alternative methodology to address many of the issues associated with standardized clinical diagnostics.