The measurement of a fibrinogen α C-chain 5.9 kDa fragment (FIC 5.9) using MALDI-TOF MS and a stable isotope-labeled peptide standard dilution.
The measurement of a fibrinogen α C-chain 5.9 kDa fragment (FIC 5.9) using MALDI-TOF MS and a stable isotope-labeled peptide standard dilution.
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DOI：
10.1016/j.cca.2011.02.029
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发表时间：
2011-05
期刊：
影响因子：
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通讯作者：
K. Sogawa;Y. Kodera;K. Noda;Yusuke Ishizuka;M. Yamada;H. Umemura;K. Maruyama;T. Tomonaga;O. Yokosuka;F. Nomura
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作者：
K. Sogawa;Y. Kodera;K. Noda;Yusuke Ishizuka;M. Yamada;H. Umemura;K. Maruyama;T. Tomonaga;O. Yokosuka;F. Nomura
背景我们之前确定了纤维蛋白原 α C 链的 5.9 kDa 肽片段（FIC 5.9）作为酗酒的新型生物标志物候选物。为了改进 FIC 5.9 测量以使其在临床诊断中的潜在用途，我们将 ClinProt 系统和稳定同位素标记的肽标准稀释液结合起来，作为测量 FIC 5.9 的简单且可重复的系统。方法我们分析了从酒精依赖患者、慢性丙型肝炎患者和健康志愿者获得的 104 份血清样本。使用 ClinProt 系统测量血清 FIC 5.9 水平，并使用或不使用稳定同位素标记的合成 FIC 5.9 作为内标。 结果稳定同位素稀释质谱 (SID-MS) 的日内和日间 CV 明显小于传统 MALDI-TOF MS。在两个不同的 MALDI-TOF MS 平台中，我们与 SID-MS 获得了一致的结果。此外，只有SID-MS检测到慢性丙型肝炎组和对照组的血清FIC 5.9水平存在微小但显着的差异。结论带有稳定同位素标记肽尖峰的MALDI-TOF MS可以比传统MS更精确地测定血清FIC 5.9水平。这将使实验室间 FIC 5.9 比较成为可能。
BackgroundWe previously identified a 5.9ákDa peptide fragment of fibrinogen α C-chain (FIC 5.9) as a novel biomarker candidate for heavy drinking. In an effort to improve FIC 5.9 measurement for potential use in clinical diagnostics, we combined the ClinProt System and a stable isotope-labeled peptide standard dilution as a simple and reproducible system for measuring FIC 5.9.MethodsWe analyzed 104 serum samples that were obtained from patients with alcohol dependency, from patients with chronic hepatitis C, and from healthy volunteers. Serum FIC 5.9 levels were measured using the ClinProt system with and without a stable isotope-labeled synthetic FIC 5.9 as an internal standard.ResultsThe within-day and between-day CVs were significantly smaller with stable isotope dilution mass spectrometry (SID-MS) than with conventional MALDI-TOF MS. Of the two different MALDI-TOF MS platforms, we obtained concordant results with SID-MS. Furthermore, only SID-MS detected a small but significant difference between the serum FIC 5.9 levels in the chronic hepatitis C group and the controls.ConclusionsMALDI-TOF MS with a stable isotope-labeled peptide spike can determine serum FIC 5.9 levels more precisely than conventional MS. This will make inter-laboratory FIC 5.9 comparisons possible.