The emergence of high-throughput
protein quantification methodologies has enabled the comprehensive characterization by longitudinal and cross-sectional studies of biological fluids under physiological and pathological conditions. In particular, the simultaneous investigation of
cytokines and
growth factors signaling pathways and their associated downstream effectors by integrated multiplexed approaches offers a powerful strategy to gain insights into biological networks and processes in living systems. A growing body of research indicates that bioactive molecules of human reproductive fluids, including human follicular fluid (hFF), may affect oocyte quality, fertilization and embryo development, thus potentially influencing the physiopathology of pregnancy-related conditions. In this work, an iTRAQ labeling strategy has been complemented with a multiplexed
protein array approach to analyze hFFs with the aim to investigate biological processes and pathways related to in vitro fertilization (IVF) outcome. The iTRAQ labeling strategy lead to the quantification of 89
proteins, 30 of which were differentially expressed in hFFs with successful compared to unsuccessful IVF outcome. The targeted study, based on multiplexed antibody
protein arrays, allowed the simultaneous quantification of 27 low abundance
proteins, including
growth factors,
chemokines and
cytokines endowed with pro- and anti-inflammatory activity. A significant number of differentially regulated
proteins were involved in biological functions related to blood coagulation,
acute phase response signaling and
complement system. Overall, the present results provide an integrated overview of
protein changes in hFFs associated to IVF outcome, thus improving current knowledge in reproductive medicine and fertility research.