Hepcidinanalysis.com is an initiative to serve the scientific and medical communities with high-quality hepcidin measurements to study the role of this peptide hormone in disorders of iron metabolism.
In 2005, we were the first to employ time-of-flight mass spectrometry to measure hepcidin in urine. We have continued to improve this assay towards a current quantitative assay to measure hepcidin in human serum and urine samples using an internal standard for quantification. In 2010, we have also developed and validated a competitive ELISA, that facilitates measurements for studies with large sample sizes. Using both assays, we have gained thorough expertise on biomedical and pre-analytical factors that influence hepcidin levels in clinical and biological samples and our assay has been used in several biomedical studies. We now coordinate the harmonization of hepcidin methods that are currently in use throughout the world.
We offer quantitative serum/plasma and urine hepcidin measurements by two different methods:
1. A combination of weak cation exchange chromatography and time-of-flight mass spectrometry (WCX-TOF MS) using a hepcidin analogue as internal standard. This method allows discrimination between the four naturally occurring isoforms, hepcidin-20, -22, -24 and -25, which are the result from differential processing at the hepcidin amino-terminus. Importantly, only the 25 amino acid form can effectively block the iron transporter ferroportin and is referred to as “bioactive” hepcidin.
2. A competitive ELISA. This method measures the total of hepcidin isoforms and is especially appropriate for large sample numbers.
First, we determined age- and sex-stratified reference ranges of serum hepcidin-25 concentration in a large, well-phenotyped, sample of the general population (n = 2998), and defined the reference [hepcidin/ferritin] and [transferrin saturation/hepcidin] ratios, which are important for the clinical interpretation of measured hepcidin concentrations. Second, reference ranges of serum hepcidin-25 concentration and [hepcidin/ferritin] ratio were determined in a population of healthy children, aged 6 months to 3 years old. Third, we defined reference ranges for serum hepcidin-25 in specified time periods during pregnancy.
Our service for clinical trials and patient care complies with both GCP regulations and with the accreditation criteria for medical laboratories as laid down in the EN/ISO 15189 and specified in the Dutch CCKL Code of practice for the implementation of a quality system. The accreditation covers the professional competence of the laboratory staff, the quality system of the laboratory as well as the specified activities and disciplines as described in the authorized annex bearing the accreditation number. We are currently working towards the NEN-EN-ISO-15189 accreditation of the Dutch Council of Accreditation.
Patient Care: hepcidin tests (human serum or plasma) can be order directly through our webform.
Clinical trials: we provide "fit-to-purpose" assay validation and subsequent analysis of samples from (pre-)clinical trials. Personnel involved in the study conduct are adequately trained in General Clinical Practice.
Collaborative projects: we assist in the development/validation of new assays and offer hepcidin measurements at a reduced fee if the focus of a project is on gaining new insights in iron metabolism and will lead to joint publications.
General Procedure Trials & Projects
1. Send an email to firstname.lastname@example.org to specify your interest in our service
2. A Service Request Form will be returned, together with answers to project-specific questions and a cost indication
3. Fill-in and return Service Request Form to email@example.com
5. Send-in sample(s) together with countersigned Project Agreement.
6. Send electronic list with sample ID’s (Excel) to firstname.lastname@example.org as shipment dispatch notification
7. Hepcidin Data Sheet and Project Invoice will be returned by email
General Sample Recommendations
Preferred use of serum/plasma and standardized sampling time; apply 0.5-1 ml fresh sample in 2 ml polypropylene tube; store at -80°C avoiding freeze-thaw cycles; if possible standardized sampling and storage protocols for cases & controls; transportation on dry ice per express mail.
Animal Models & Other Applications
We have established assay for mouse hepcidin peptides in serum/plasma (Hep-1) and urine (Hep-2), for rat hepcidin in urine, and for monkey, pig and cow serum/plasma hepcidin-25. Our flexible WCX-TOF MS approach also allows hepcidin measurements in other human body fluids and growth medium from in vitro cultured human cell lines.
Hepcidinanalysis.com is located within the Department of Laboratory Medicine of the Radboud University Nijmegen Medical Center and is embedded within the Institute for Genetic and Metabolic Diseases. The team members have a broad expertise in iron metabolism, clinical chemistry, diagnostic medicine, biomedical sciences, molecular biology, mass spectrometry and biochemistry.
Hepcidinanalysis.com is part of the legal entity “UMC St Radboud Onderzoek Klinische Chemie B.V.”, which is listed in the Commercial Register of the Chamber of Commerce under file number 41261285.