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

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


  1. Procalcitonin (PCT)-guided antibiotic stewardship: an international experts consensus on optimized clinical use

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Individual values of antineutrophil cytoplasmic antibodies do not correspond between antigen-specific assays

    Research output: Contribution to journalLetterResearchpeer-review

  1. Udredning og behandling af mandlig infertilitet

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. DIAGNOSIS OF ENDOCRINE DISEASE: Sex steroid action in adolescence: too much, too little; too early, too late

    Research output: Contribution to journalReviewResearchpeer-review

  3. Familial resemblance in markers of testicular function in fathers and their young sons: a cross-sectional study

    Research output: Contribution to journalJournal articleResearchpeer-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
Issue number11
Pages (from-to)1913-20
Publication statusPublished - 2018

ID: 54666727