Research
Print page Print page
Switch language
Rigshospitalet - a part of Copenhagen University Hospital
Published

Development and validation of a mass spectrometry-based assay for quantification of insulin-like factor 3 in human serum

Research output: Contribution to journalJournal articleResearchpeer-review

DOI

  1. High incidence of discrepancies in new Siemens assay - A comparison of cardiac troponin I assays

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Coagulation parameters in the newborn and infant - the Copenhagen Baby Heart and COMPARE studies

    Research output: Contribution to journalJournal articleResearchpeer-review

  1. Dynamic changes of reproductive hormones in male minipuberty: Temporal dissociation of Leydig- and Sertoli-cell activity

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Oophorectomy and rate of dementia: a prospective cohort study

    Research output: Contribution to journalJournal articleResearchpeer-review

  3. Pregnancy exposure to bisphenol A and duration of breastfeeding

    Research output: Contribution to journalJournal articleResearchpeer-review

  4. Environmental factors in declining human fertility

    Research output: Contribution to journalReviewResearchpeer-review

View graph of relations

BACKGROUND: The circulating level of the peptide hormone insulin-like factor 3 (INSL3) is a promising diagnostic marker reflecting Leydig cell function in the male. Few commercial immunoassays of varying quality exist. Therefore, we decided to develop and validate a precise method for quantification of INSL3 by mass spectrometry.

METHODS: We developed an assay in which the INSL3 A-chain is released from the INSL3 A-B heterodimer by chemical reduction and alkylation. The alkylated INSL3 A-chain is quantitated by liquid chromatography-tandem mass spectrometry (LC-MS/MS), as substitute for serum INSL3. The method was compared to a validated and sensitive in-house serum INSL3 immunoassay using 97 serum samples from 12 healthy boys during pubertal transition. Adult levels were determined based on sera from 72 adult healthy males aged 18-40 years.

RESULTS: An LC-MS/MS assay with limit of detection and limit of quantification (LOQ) of 0.06 and 0.15 ng/mL, respectively, and intra-assay CVs <9% in the relevant ranges was obtained. The LC-MS/MS compared well with the in-house immunoassay (Deming regression slope: 1.28; Pearson correlation: R=0.86). INSL3 concentrations increased with pubertal maturation in healthy boys. INSL3 concentrations were above the LOQ in all samples from the adult men. The mean (±2 SD range)for serum INSL3 concentrations in the adult men was 2.2 (0.5-3.9) ng/mL.

CONCLUSIONS: We have developed a robust and sensitive method suitable for quantitation of serum INSL3 in a clinical setting using LC-MS/MS instrumentation available in modern clinical laboratories. The method paves the way for future studies into the clinical role of serum INSL3 measurements.

Original languageEnglish
JournalClinical Chemistry and Laboratory Medicine
Volume56
Issue number11
Pages (from-to)1913-20
ISSN1434-6621
DOIs
Publication statusPublished - 2018

ID: 54666727