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The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis

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Abstract

A noninvasive diagnostic test for endometriosis is needed to shorten the current diagnostic delay of 8–11 years. The goal of this study was to discover new biomarkers for endometriosis using an antibody array approach. A total of 103 plasma samples from patients with laparoscopically confirmed presence (n = 68) or absence (n = 35) of endometriosis were selected. Samples were pooled according to disease status, cycle phase, disease stage, and phenotype. Pooled samples were screened for possible biomarkers using the L-series 1000 and Quantibody 660 arrays from RayBiotech. Technical verification of ten markers was done using a custom-made multiplex immunoassay identifying ten proteins (10-plex) and later by single ELISA. Due to the limited reproducibility of the L-series 1000 immunoassay, the biomarker screening was performed using the Quantibody 660, a sandwich-based multiplex immunoassay, which showed that 280 proteins were upregulated, and 29 proteins downregulated in the endometriosis pool versus the control pool. In order to assess the reproducibility of these results, ten preselected proteins were analyzed using a custom 10-plex. Four proteins (CD48, DNAM-1, IL-31, and XIAP) were confirmed to be differentially expressed when comparing the endometriosis and control pool. However, only IL-31 showed a univariate statistical difference between endometriosis and control groups in individual samples that were part of the initial pools. In conclusion, discovery and verification of potential markers proved challenging using multiplex immunoassay methods, mainly due to issues with reproducibility. Only IL-31 showed potential as possible biomarker for endometriosis.

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Acknowledgments

We thank Ana Arraztio (TebuBio) and the staff of RayBiotech, in particular Jarad Wilson (RayBiotech), for excellent technical support; M. Welkenhuysen for acquiring patient consent; G. Vriens, C. Tomassetti, C. Meuleman and K. Peeraer for their contributions to the sample collection; all patients who have given consent for sample collection.

Funding

Grant support: Funding of this research was obtained from the Research Foundation-Flanders (FWO, application number 11X5515N).

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Authors and Affiliations

  1. Department of Development and Regeneration, Woman and Child, KU Leuven, Leuven, Belgium

    Dorien O, Arne Vanhie, Daniëlle Peterse, Amelie Fassbender & Thomas D’Hooghe

  2. Facility for systems biology based mass spectrometry, KU Leuven, Leuven, Belgium

    Etienne Waelkens

  3. Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium

    Etienne Waelkens

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  1. Dorien O
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  6. Thomas D’Hooghe
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Correspondence to Thomas D’Hooghe.

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Conflict of Interest

Prof Dr. Thomas D’Hooghe is Vice President and Head Global Medical Affairs Fertility, Research and Development, Merck KGaA, Darmstadt, Germany. He is also a Professor in Reproductive Medicine and Biology at the Department of Development and Regeneration, Group Biomedical Sciences, KU Leuven (University of Leuven), Belgium, and an Adjunct Professor at the Department of Obstetrics and Gynecology in the University of Yale, New Haven, USA. Neither his corporate role nor his academic roles represent a conflict of interest with respect to the work done by him for this study.

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Amelie Fassbender Thomas D’Hooghe should be regarded as joint last authors

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O, D., Waelkens, E., Vanhie, A. et al. The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis. Reprod. Sci. 27, 751–762 (2020). https://doi.org/10.1007/s43032-019-00081-w

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  • DOI: https://doi.org/10.1007/s43032-019-00081-w

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