Anti-Müllerian hormone (AMH) is a biomarker for the assessment of female fertility. The accurate measurement of the concentration of AMH is relevant for the success of assisted reproductive therapies and diagnosis of clinical cases. In this study, we show that cytokines such as fetal liver tyrosine kinase 3 ligand (Flt3L), CC subtype chemokine ligand 20 (CCL20), granulocyte–macrophage colony-stimulating factor (GM-CSF), and β2-microglobulin (β2M) significantly enhance the immune response against AMH. Two anti-AMH monoclonal antibodies (mAbs) with high affinity were selected by biolayer interferometry (BLI) technology for application in a fully automated magnetic chemiluminescence immunoassay (CLIA). This robust and rapid assay can efficiently detect AMH in the range of 0.125~20 ng mL−1 with a detection limit of 0.099 ng mL−1. This immunoassay showed high specificity with no cross-reaction with structurally related proteins and some of the other members of the TGF-β super family, such as inhibin A, activin A, follicle-stimulating hormone, and luteinizing hormone. The average recovery rates of three different batches were 100.19%, 102.72%, and 103.59%, respectively, with coefficients of variation of less than 12%. The developed assay was applied in the detection of AMH in 69 serum samples from randomly selected patients. Our data showed a high correlation with those obtained using commercially available ELISA kits (correlation coefficient, 0.9831). Hence, we suggest that this immunoassay could find application in the development of POCT for the diagnosis of AMH in clinical samples.
Anti-Müllerian hormone Cytokine DNA immunization High-affinity antibodies Point-of-care testing
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This work is supported by the Program of Guangzhou Science & Technology (201802010030).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All animal experiments and welfare of the animals were performed under ethical approval from and in agreement with the guidelines of the Institutional Animal Care and Use Ethics Committee of South China University of Technology and also in accordance with the policy of the National Ministry of Health. Written informed consents were obtained from all individual participants who provided the serum samples.
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