Immunochemical and Mass-Spectrometry-Based Serum Hepcidin Assays for Iron Metabolism Disorders
Immunochemical and Mass-Spectrometry-Based Serum Hepcidin Assays for Iron Metabolism Disorders
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DOI：
10.1373/clinchem.2010.149187
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发表时间：
2010-10-01
影响因子：
9.3
通讯作者：
Swinkels, Dorine W.
中科院分区：
文献类型：
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作者：
Kroot, Joyce J. C.;Laarakkers, Coby M. M.;Swinkels, Dorine W.
背景：海普西丁是一种铁调节多肽激素，由3种亚型组成：生物活性海普西丁-25、非活性海普西丁-22和海普西丁-20。海普西丁用于铁代谢紊乱的诊断和监测，但其在血清中的可靠定量方法很少，对其相对分析强度和临床应用的了解也很少。方法：我们建立了竞争性(C)-ELISA和免疫捕获TOF-MS(IC-TOF-MS)分析方法。利用这两种方法和我们先前描述的弱阳离子交换(WCX)-TOF-MS方法，我们测量了186例各种铁代谢紊乱患者和23名健康对照的血清中庚西丁浓度。结果：我们发现：(A)不同疾病中的中位数肝西汀浓度的相对差异是相似的，尽管c-ELISA和WCX-TOF-MS的绝对浓度不同；(B)不同方法之间的肝西汀浓度的差异，在慢性肾脏疾病患者中最为显著；以及(C)c-ELISA和IC-TOF-MS测定的肝西汀浓度与铁蛋白浓度相关
BACKGROUND: Hepcidin is an iron-regulatory peptide hormone that consists of 3 isoforms: bioactive hepcidin-25, and inactive hepcidin-22 and hepcidin-20. Hepcidin is instrumental in the diagnosis and monitoring of iron metabolism disorders, but reliable methods for its quantification in serum are sparse, as is knowledge of their relative analytical strengths and clinical utility.METHODS: We developed a competitive (c)-ELISA and an immunocapture TOF mass-spectrometry (IC-TOF-MS) assay. Exploiting these 2 methods and our previously described weak cation exchange (WCX)-TOF-MS assay, we measured serum hepcidin concentrations in 186 patients with various disorders of iron metabolism and in 23 healthy controls.RESULTS: We found that (a) the relative differences in median hepcidin concentrations in various diseases to be similar, although the absolute concentrations measured with c-ELISA and WCX-TOF-MS differed; (b) hepcidin isoforms contributed to differences in hepcidin concentrations between methods, which were most prominent in patients with chronic kidney disease; and (c) hepcidin concentrations measured by both the c-ELISA and IC-TOF-MS correlated with ferritin concentrations