Home » Abstracts » Human Health Proteomics » Single human oocyte analysis by LC-MS/MS. Approach validation and characterization of endometriosis patients’ oocytes.

Single human oocyte analysis by LC-MS/MS. Approach validation and characterization of endometriosis patients’ oocytes.

Human Health Proteomics

Abstract #270 Posted on28 March 202512 May 2025 12 May 2025 12h22

Abstract

In Vitro Fertilization (IVF) is a key assisted reproductive technology (ART) in which human oocytes are fertilized with sperm in a laboratory setting. Although this approach is one of the most effective ART methods, most in vitro-produced embryos fail to implant. Oocyte quality may be one of the key limiting factors in IVF success. In this context, current proteomic techniques can provide valuable insights into the characterization of human oocyte protein content.

In this study, we characterized a set of 71 single human oocytes, including Germinal Vesicle (GV), Metaphase I (MI), and Metaphase II (MII) stages, from women with and without endometriosis. Samples were prepared using ultrasensitive paramagnetic bead technology (SP3) and analyzed on an LC-MS/MS system comprising an Evosep ONE (Evosep) liquid chromatograph and a tims TOF Pro (Bruker) mass spectrometer. Data were acquired in Data-Independent Acquisition (DIA) mode and analyzed using DIA-NN v1.8.1.

Our approach enabled the reproducible identification and quantification of more than 1,700 proteins. The consistent variation of bona fide maturation-related proteins through different oocyte stages (PCNA, YBOX1, WEE2) validated our approach. Additionally, we compared the proteomic profile of oocytes from endometriosis patients and women without endometriosis. These comparisons revealed specific protein expression signatures in endometriosis that should are currently being further characterized.

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In this study, we characterized a set of 71 single human oocytes, including Germinal Vesicle (GV), Metaphase I (MI), and Metaphase II (MII) stages, from women with and without endometriosis. Samples were prepared using ultrasensitive paramagnetic bead technology (SP3) and analyzed on an LC-MS/MS system comprising an Evosep ONE (Evosep) liquid chromatograph and a tims TOF Pro (Bruker) mass spectrometer. Data were acquired in Data-Independent Acquisition (DIA) mode and analyzed using DIA-NN v1.8.1.

Our approach enabled the reproducible identification and quantification of more than 1,700 proteins. The consistent variation of bona fide maturation-related proteins through different oocyte stages (PCNA, YBOX1, WEE2) validated our approach. Additionally, we compared the proteomic profile of oocytes from endometriosis patients and women without endometriosis. These comparisons revealed specific protein expression signatures in endometriosis that should are currently being further characterized.

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