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

Anand-Ivell, Ravinder; Frederiksen, Hanne; Juul, Anders; Andersson, Anna-Maria; Albrethsen, Jacob; Nordkap, Loa; Kirstine Bang, Anne; Jørgensen, Niels

Authors

Ravinder Anand-Ivell ravinder.anand-ivell@nottingham.ac.uk

Hanne Frederiksen

Anders Juul anders.juul@regionh.dk

Anna-Maria Andersson

Jacob Albrethsen Jakob.Christian.Albrethsen@regionh.dk

Loa Nordkap

Anne Kirstine Bang

Niels Jørgensen



Abstract

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

Journal Article Type Article
Publication Date May 30, 2018
Journal Clinical Chemistry and Laboratory Medicine
Print ISSN 1434-6621
Electronic ISSN 1437-4331
Publisher De Gruyter
Peer Reviewed Peer Reviewed
Volume 56
Issue 11
APA6 Citation Anand-Ivell, R., Juul, A., & Albrethsen, J. (2018). Development and validation of a mass spectrometry-based assay for quantification of insulin-like factor 3 in human serum. Clinical Chemistry and Laboratory Medicine, 56(11), doi:10.1515/cclm-2018-0171
DOI https://doi.org/10.1515/cclm-2018-0171
Keywords assay development; insulin-like factor 3 (INSL3); mass spectrometry
Publisher URL https://www.degruyter.com/view/j/cclm.2018.56.issue-11/cclm-2018-0171/cclm-2018-0171.xml
Copyright Statement Copyright information regarding this work can be found at the following address: http://eprints.nottingh.../end_user_agreement.pdf
Additional Information

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Files

INSL3 method paper LCMS 2018.pdf (638 Kb)
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Copyright Statement
Copyright information regarding this work can be found at the following address: http://eprints.nottingham.ac.uk/end_user_agreement.pdf





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