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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

Hanne Frederiksen

Anders Juul

Anna-Maria Andersson

Jacob Albrethsen

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

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

Journal Article Type Article
Acceptance Date May 3, 2018
Online Publication Date May 30, 2018
Publication Date May 30, 2018
Deposit Date Jun 1, 2018
Publicly Available Date May 31, 2019
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
DOI https://doi.org/10.1515/cclm-2018-0171
Keywords assay development; insulin-like factor 3 (INSL3); mass spectrometry
Public URL http://eprints.nottingham.ac.uk/id/eprint/52150
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.nottingham.ac.uk/end_user_agreement.pdf
Additional Information

Composition Type:

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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