High-sensitive radioimmunoassay for human serum hepcidin.
The hepatic peptide hormone hepcidin plays a central role in body iron metabolism. Despite its promise as a biomarker, the availability of high-sensitive hepcidin assays is still limited. We developed and validated a RadioImmunoAssay (RIA) to measure hepcidin quantitatively in human serum. This assay exhibited a very low detection limit (0.02 microg/l), low imprecision (coefficient of variation-range 4.4-6.2%) and good linearity and recovery (range: 81-105%). Hepcidin levels of samples of controls and patients with iron deficiency and inflammation showed an excellent correlation with our previously described quantitative time-of-flight mass spectrometry assay (range 2.5-266.8 microg/l, r = 0.92, P < 0.0001). The RIA detected: (i) differences in mean hepcidin levels between men (n = 29) and women (n = 35), (ii) differences between individuals of different HFE-genotypes (n = 60) and (iii) daily increases in hepcidin levels (n = 64). The assay (i) is easy to perform and many samples can be processed within one assay-run, (ii) shows accurate, reproducible and high-sensitive measurements and (iii) is anticipated to be particularly useful to study the effects of pathological and physiological stimuli on hepcidin levels in the lower range.
Grebenchtchikov
Nicolai
N
Department of Chemical Endocrinology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
Geurts-Moespot
Anneke J
AJ
Kroot
Joyce J C
JJ
den Heijer
Martin
M
Tjalsma
Harold
H
Swinkels
Dorine W
DW
Sweep
Fred G J
FG
D000328:Adult; D018798:Anemia, Iron-Deficiency / Q000097:blood; D023181:Antimicrobial Cationic Peptides / Q000097:blood*; D015415:Biomarkers / Q000097:blood; D002940:Circadian Rhythm; D005260:Female; D006432:Hemochromatosis / Q000097:blood; D064451:Hepcidins; D006801:Humans; D007249:Inflammation / Q000097:blood; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D011863:Radioimmunoassay / Q000379:methods*; D012016:Reference Values; D012680:Sensitivity and Specificity; D055815:Young Adult
D023181:Antimicrobial Cationic Peptides; D015415:Biomarkers; C578702:HAMP protein, human; D064451:Hepcidins