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High-Throughput Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry Method for Quantification of Hepcidin in Human Urine

机译:高通量基质辅助激光解吸电离-飞行时间质谱法定量人尿中的铁调素

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摘要

Levels of the peptide hormone hepcidin negatively correlatenwith systemic iron status and are increased inndisorders in which iron metabolism is secondarily disregulated,nsuch as the anemia of chronic disease. Consequently,nthe ability to measure hepcidin in the clinicalnsetting may have diagnostic value for a broad range ofnindications. We describe a novel quantitative matrixassistednlaser desorption ionization-time-of-flight (MALDITOF)nmass spectrometry assay for hepcidin in humannurine which involves (i) direct enrichment from minutenvolumes (5 μL) of minimally treated urine on the surfacenof a functionalized chip, (ii) quantification by the use of anstable isotope labeled internal standard, and (iii) analysisnby MALDI-TOF. Performance features include a widenlinear range (1-1000 nM; LOQ 2.5 nM), high accuracyn(90-110% recovery) and precision (intraday CV 12.11%;ninterday CV 13.21%), and a strong correlation uponninterlaboratory cross validation with an existing immunoassay.nThe assay is simple, accurate, and efficient, andnthe high-throughput performance features of the assaynmake large-scale clinical research studies feasible.
机译:肽激素铁调素的水平与全身铁状态负相关,并且是铁代谢继而失调的疾病,例如慢性疾病的贫血。因此,在临床环境中测量铁调素的能力可能对广泛的适应症具有诊断价值。我们描述了一种新型定量基质辅助激光解吸电离飞行时间(MALDITOF)nmass光谱测定人体中的hepcidin的方法,其中涉及(i)从功能化芯片表面上经过最小处理的每分钟体积(5μL)尿液中直接富集,(ii )通过使用不稳定的同位素标记的内标进行定量,以及(iii)通过MALDI-TOF进行分析。性能特点包括宽线性范围(1-1000 nM; LOQ 2.5 nM),高精度n(90-110%回收率)和精度(日内CV 12.11%;日间CV 13.21%),以及实验室间交叉验证与现有产品之间的强相关性免疫测定。该测定简单,准确,高效,并且测定的高通量性能使大规模临床研究可行。

著录项

  • 来源
    《Analytical Chemistry》 |2010年第4期|p.1551-1555|共5页
  • 作者单位

    Departments of Pathology and Medicine, Children’s Hospital Boston, Boston, Massachusetts 02115, Departments ofPathology and Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, Proteomics Center at Children’sHospital Boston, Boston, Massachusetts 02115, and QIAGEN GmbH, Hilden, Germany 40724;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
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  • 入库时间 2022-08-17 13:36:33

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