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

Clinical Chemistry and Laboratory Medicine : CCLM
Jakob AlbrethsenAnders Juul

Abstract

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. 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. 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 ...Continue Reading

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Citations

Mar 27, 2020·The Journal of Clinical Endocrinology and Metabolism·Jakob AlbrethsenAnders Juul
Sep 29, 2020·Frontiers in Endocrinology·Jakob AlbrethsenAnna-Maria Andersson
Aug 8, 2020·The Journal of Clinical Endocrinology and Metabolism·Jakob AlbrethsenAnders Juul
Oct 16, 2019·Frontiers in Endocrinology·Steven M HarrisonLinda A Baker
Sep 9, 2020·The Journal of Clinical Endocrinology and Metabolism·Jorma Toppari
Mar 15, 2021·Andrology·Ravinder Anand-IvellRichard Ivell
Aug 18, 2020·General and Comparative Endocrinology·Richard IvellRavinder Anand-Ivell
Mar 12, 2021·The Journal of Clinical Endocrinology and Metabolism·Jon Jarløv RasmussenCaroline Kistorp
Sep 29, 2020·Protein and Peptide Letters·Paolo FacondoAlberto Ferlin
Aug 28, 2021·Frontiers in Endocrinology·Marie Lindhardt LjubicicTrine Holm Johannsen

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