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The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis.
Reprod Sci. 2020 02; 27(2):751-762.RS

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.

Authors+Show Affiliations

Department of Development and Regeneration, Woman and Child, KU Leuven, Leuven, Belgium.Facility for systems biology based mass spectrometry, KU Leuven, Leuven, Belgium. Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium.Department of Development and Regeneration, Woman and Child, KU Leuven, Leuven, Belgium.Department of Development and Regeneration, Woman and Child, KU Leuven, Leuven, Belgium.Department of Development and Regeneration, Woman and Child, KU Leuven, Leuven, Belgium.Department of Development and Regeneration, Woman and Child, KU Leuven, Leuven, Belgium. Thomas.dhooghe@kuleuven.be.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32016799

Citation

O, Dorien, et al. "The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis." Reproductive Sciences (Thousand Oaks, Calif.), vol. 27, no. 2, 2020, pp. 751-762.
O D, Waelkens E, Vanhie A, et al. The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis. Reprod Sci. 2020;27(2):751-762.
O, D., Waelkens, E., Vanhie, A., Peterse, D., Fassbender, A., & D'Hooghe, T. (2020). The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis. Reproductive Sciences (Thousand Oaks, Calif.), 27(2), 751-762. https://doi.org/10.1007/s43032-019-00081-w
O D, et al. The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis. Reprod Sci. 2020;27(2):751-762. PubMed PMID: 32016799.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The Use of Antibody Arrays in the Discovery of New Plasma Biomarkers for Endometriosis. AU - O,Dorien, AU - Waelkens,Etienne, AU - Vanhie,Arne, AU - Peterse,Daniëlle, AU - Fassbender,Amelie, AU - D'Hooghe,Thomas, Y1 - 2020/02/03/ PY - 2019/04/18/received PY - 2019/07/05/accepted PY - 2020/2/6/pubmed PY - 2020/2/6/medline PY - 2020/2/5/entrez KW - Antibody array KW - Biomarker KW - Endometriosis KW - Immunoassay KW - Reproducibility SP - 751 EP - 762 JF - Reproductive sciences (Thousand Oaks, Calif.) JO - Reprod Sci VL - 27 IS - 2 N2 - 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. SN - 1933-7205 UR - https://www.unboundmedicine.com/medline/citation/32016799/The_Use_of_Antibody_Arrays_in_the_Discovery_of_New_Plasma_Biomarkers_for_Endometriosis_ L2 - https://journals.sagepub.com/doi/10.1007/s43032-019-00081-w?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -
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