Updated publication reference for PubMed record(s): 31223520. Background. Silkworm pupae (SWP) is the main by-product of the sericulture industry with an interesting nutritional profile, especially in terms of proteins. In consideration of its possible use as a food or food ingredient in Western countries, a comparative proteomic experiment has been performed to investigate the differences of the protein profile of male and female SWP reared on mulberry leaves or on an artificial diet. Methods. The nutritional profile of lyophilized SWP in terms of dry matter and ash was evaluated according to the AOAC procedures, the total nitrogen content was determined by a nitrogen analyzer and the SWP gross energy value was measured using an adiabatic calorimetric bomb. The comparative proteomic analysis was performed on male and female SWP reared on mulberry leaves or on the artificial diet. Proteins were separated by Bidimensional Electrophoresis (2DE) and, after a multivariate statistical analysis, the differentially expressed proteins were identified by LC-MS/MS. Results. The comparative proteomic approach highlighted 47 SWP proteins differentially expressed comparing diet and gender. PCA analysis showed that 7 proteins were more effective in discriminating the sex and 5 were more effective in discriminating the diet type. In spite of the above mentioned differences in the SWP protein profile, no strong alteration of the pupa physiological traits have been demonstrated, suggesting a general SWP flexibility to adapt to a well-balanced artificial diet. Differences in lipid transport and metabolism were found among the experimental groups, that might have a relevant effect on the timing, on hormone secretion and, in turn, on insect voltinism. This aspect may also affect silk production, as univoltine strains are the most productive. Although this is a preliminary study, the proteomic data may offer a contribution in understanding also the influence of gender and farming strategy on the allergen profile of B. mori, when used as food or as a food ingredient. Since female silkworm pupae reared on mulberry leaves seemed to contain lower levels of known allergens than those reared in the other experimental conditions, we speculated that these findings may help when farming B. mori for food production purposes. However, these results need to be supported by further characterization of the allergenic potential of B. mori.