Egg quality is of pivotal importance in biomedicine, agriculture, ecology and environmental science because of its tremendous influence on reproductive success or failure in all animals. Despite species specific differences in physiological aspects of early development, the evolutionary conserved stereotypical procedure of cellular events, led us to investigate whether these findings are common with marine fish species of aquaculture interest. Atlantic halibut (Hippoglossus hippoglossus) is a highly prized species in global fish markets with increasing demand to its production. The objectives of this study were 1) to reveal the proteomic profiles of good versus poor quality halibut eggs, 2) to identify proteins that can serve as egg quality markers, and 3) to discover molecular mechanisms determining egg quality using a combination of quantitative proteomics workflows, tandem mass tags (TMT) labeling and parallel reaction monitoring (PRM).