SELDI-TOF-MS determination of hepcidin in clinical samples using stable isotope labelled hepcidin as an internal standard

Ward, Douglas G.; Roberts, Keith; Stonelake, Paul S.; Goon, Patrick K. Y.; Zampronio, Cleidiane G.; Martin, Ashley; Johnson, Philip J.; Iqbal, Tariq; Tselepis, Chris

doi:10.1186/1477-5956-6-28

Cited by 61 publications

(57 citation statements)

References 28 publications

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“…In addition, Ganz et al (29 ) developed a competitive (c)ELISA for human serum hepcidin. Other groups have also reported reliable hepcidin assays that can be divided in 3 main methodologies: (a) MS (85)(86)(87)(88)(89)(90)(91)(92)(93)(94)(95); (b) immunochemical assays, comprising cRIA (96,97 ), cELISA (13,29,98,99 ), and a 2-site ELISA (100 ); and (c) a ligand-binding assay (43 ) ( Table 1). Of the currently available commercial immunochemical research kits for serum hepcidin, we found the RIA and enzyme-immunoassay kits of Bachem (purchased November 2009 and August 2010, respectively) to be suitable to differentiate between hepcidin concentrations in serum samples of controls and patients with various iron disorders, whereas the bioactive hepcidin kit of DRG Instruments, (purchased October 2009) gave similar concentrations for all samples and could not discriminate between iron metabolism disorders (unpublished results).…”

Section: Second-generation (Quantitative) Hepcidin Assaysmentioning

confidence: 99%

Hepcidin in Human Iron Disorders: Diagnostic Implications

et al. 2011

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BACKGROUND:The peptide hormone hepcidin plays a central role in regulating dietary iron absorption and body iron distribution. Many human diseases are associated with alterations in hepcidin concentrations. The measurement of hepcidin in biological fluids is therefore a promising tool in the diagnosis and management of medical conditions in which iron metabolism is affected.CONTENT: We describe hepcidin structure, kinetics, function, and regulation. We moreover explore the therapeutic potential for modulating hepcidin expression and the diagnostic potential for hepcidin measurements in clinical practice.

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Section: Second-generation (Quantitative) Hepcidin Assaysmentioning

confidence: 99%

Hepcidin in Human Iron Disorders: Diagnostic Implications

et al. 2011

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“…Recently, Ward et al (34) used an isotopically labeled hepcidin as an internal standard for the quantitation of hepcidin using SELDI-TOF MS. No accuracy and recovery data have been presented; however, there is good intraday (Ͻ10%) and interday (Ͻ20%) precision with this assay. The lowest quantity of internal standard reliably detected in serum was 10 ng/ml, and this study found the average hepcidin concentration in healthy women to be 50 ng/ml.…”

Section: Surface Enhanced Laser Desorption/matrix-assisted Laser Desomentioning

confidence: 99%

Current Status of the Measurement of Blood Hepcidin Levels in Chronic Kidney Disease

Macdougall¹,

Małyszko²,

Hider³

et al. 2010

Clinical Journal of the American Society of Nephrology

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Hepcidin is a small defensin-like peptide produced in the liver in response to anemia, hypoxia, or inflammation. In addition to its anti-microbial properties, it has also been found to be a key regulator of iron utilization, providing increased understanding of why chronic kidney disease patients absorb iron poorly from the gut and also why many hemodialysis patients develop functional iron deficiency in the presence of inflammation. Hepcidin synthesis is upregulated in uremia, as in other inflammatory states. The ability to measure hepcidin in biologic fluids has stimulated interest in the potential applicability of this measurement as a more informative marker of iron status than the traditional iron indices such as serum ferritin and transferrin saturation. Until recently, however, the assays for measuring hepcidin have lacked precision, accuracy, and internal validation. Over the last few years, however, several assays have become available that address these limitations. Broadly speaking, these can be divided into radioimmunoassays, ELISAs, and mass spectrometry methods. The purpose of this review is to outline the various assays available at the present time, to critique their advantages and limitations, and to report comparative data in patients with chronic kidney disease. A concern with the immunoassays is that they detect more than biologically active hepcidin-25. Mass spectrometric assays are specific for hepcidin-25 but are labor intensive and require more costly and sophisticated instrumentation. Thus, although mass spectrometry is more accurate, it is less practical for routine clinical use at the present time.

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“…Some methods use an internal standard, either hepcidin analogs or bioactive hepcidin-25 synthesized with stable isotopes. [3][4][5][6][7][8][9][10] Recently, immunochemical (IC) assays for hepcidin-25 have also been developed, which comprise of competitive radioimmunoassays (RIA) 11 and enzyme-linked immunosorbent assays (ELISA). 12,13 Currently there is no reference method for hepcidin measurements.…”

Section: Introductionmentioning

confidence: 99%