An immunoaffinity liquid chromatography-tandem mass spectrometry assay for the quantitation of matrix metalloproteinase 9 in mouse serum
An immunoaffinity liquid chromatography-tandem mass spectrometry assay for the quantitation of matrix metalloproteinase 9 in mouse serum
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DOI：
10.1016/j.ab.2010.01.002
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发表时间：
2010-04-15
影响因子：
2.9
通讯作者：
Neubert, Hendrik
中科院分区：
文献类型：
--
作者：
Ocana, Mireia Fernandez;Neubert, Hendrik
开发了一种免疫亲和液相色谱-串联质谱 (LC-MS/MS) 方法，用于定量小鼠血清中的锌内肽酶基质金属蛋白酶 9 (MMP-9)。该测定的样品制备包括使用 MMP-9 抗体进行基于磁珠的富集，并使用液体处理机器人平台以 96 孔板形式进行。通过胰蛋白酶消化衍生自 MMP-9 的替代肽 GSPLQGPFLTAR 在纳流色谱和纳喷雾电离以及选择反应监测 (SRM) 检测之前，使用包含一系列捕获柱（Cl 8、强阳离子交换和另一个 C18）的在线毛细管流动捕获释放色谱装置进行监测。该检测方法经过验证，结果准确（
An immunoaffinity liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the quantitation of the zinc endopeptidase matrix metalloproteinase 9 (MMP-9) from mouse serum. Sample preparation for the assay included magnetic bead-based enrichment using an MMP-9 antibody and was performed in a 96-well plate format using a liquid-handling robotic platform. The surrogate peptide GSPLQGPFLTAR derived from MMP-9 by trypsin digestion was monitored using an on-line capillary flow trap-release chromatography setup incorporating a series of trap columns (Cl 8, strong cation exchange, and another C18) prior to nanoflow chromatography and nanospray ionization with selected reaction monitoring (SRM) detection. The assay was fit-for-purpose validated and found to be accurate (