Quantification of Müllerian Inhibiting Substance/Anti-Müllerian Hormone polypeptide by isotope dilution mass spectrometry

• Published in: Analytical biochemistry Vol. 560; pp. 50 - 55
• Main Authors: Whiting, Gail, Ferguson, Jackie, Fang, Min, Pepin, David, Donahoe, Patricia, Matejtschuk, Paul, Burns, Chris, Wheeler, Jun X.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-11-2018
• Subjects: Anti-Müllerian hormone; Infertility; Mass spectrometry; Müllerian inhibiting substance; Selected reaction monitoring; Signature peptide; PBS; AAA; ABC; Müllerian inhibiting substance; IDMS; MS; PTM; MRM; IS; SRM; SDS-PAGE; Anti-Müllerian hormone; Infertility; MIS; SI; FWHM; Signature peptide; ESI; Selected reaction monitoring; AMH; Mass spectrometry; ELISA
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2018.05.006

Measurement of serum concentrations of Müllerian inhibiting substance (MIS), also known as anti-Müllerian Hormone (AMH) by immunoassay is gaining clinical acceptance and widespread use for the diagnosis of ovarian conditions and for prediction of the response to ovarian stimulation protocols as part of assisted reproductive therapies. Provision of an International Standard to harmonize immunoassay methods is required. It is desirable for the content of a future International Standard to be assigned in mass units for consistency with the units reported by current methods. Isotope dilution mass spectrometry (IDMS), a physicochemical method with traceability to the SI (Système International d'Unités) unit of mass, is a candidate approach to provide orthogonal data to support this mass assignment.
Here, we report on the development of an IDMS method for quantitation of AMH using three peptides from different regions of the AMH monomer as surrogates for the measurement of AMH. We show the sensitivity and linearity of the standard peptides and demonstrate the reproducibility and consistency of the measurement amongst the three peptides for determining the AMH content in buffered preparations and in trial preparations of recombinant AMH, lyophilised in the presence of an excess of bovine casein.
•Immunoassays are used to measure serum levels of AMH for the diagnosis of a number of reproductive conditions.•AMH International Standard is required for harmonization of immunoassays.•It is desirable for the content of a future International Standard to be assigned in mass units.•Isotope dilution mass spectrometry (IDMS) can be used to support mass assignment.