Investigation of a Potential Screening Tool That Uses Targeted Mass Spectrometry to Detect the Presence of Apoptotic Proteins in Conditioned Media From IVF Embryos [ID 1064]

INTRODUCTION: Assisted reproductive technologies (ARTs) are used in the treatment of infertility. One type of ART is in vitro fertilization (IVF) and according to the Centers for Disease Control and Prevention, IVF success rate was approximately 27% in 2021. This success rate is due to many factors including the mother’s age and morphology and ploidy status of the implanted embryo. This study aims to enhance IVF embryo implantation by identifying apoptotic proteins in blastocoel fluid-conditioned media from aneuploid and euploid IVF embryos using mass spectrometry. METHODS: Blastocoel fluid-conditioned media was collected from day 5 IVF embryos after preimplantation genetic testing for aneuploidy. Samples were pooled based on ploidy status. Albumin in samples was removed prior to protein concentration. In-solution trypsin digestion was performed prior to separation via Dionex UltiMate 3000 nano-flow liquid chromatograph using a C-18 reverse phase capillary column and a standard acetonitrile/water gradient. The column is interfaced to a Thermo Q-Exactive quadrupole-orbitrap mass spectrometer by an integral nanospray tip. Analysis of peptides was done by tandem mass spectrometry (LC-MS/MS) using high resolution of the fragment ions. RESULTS: Preliminary mass spectrometry of samples identified the TEK/TIE2 protein, unique to media from euploid embryos and absent in media from aneuploid embryos, along with peptides from 19 different proteins. CONCLUSIONS/IMPLICATIONS: Using mass spectrometry, this study reports the preliminary identification of a receptor tyrosine kinase, TEK/TIE2, that plays a role in the development of the vascular system. Ongoing studies are utilizing targeted mass spectrometry to assess proteins in media from euploid embryos with known implantation status to identify potential biomarkers of implantation outcomes.