Protein-based materials of animal origin, such as glue or egg, are known to have been used as binding media in paints and adhesives in ancient Egypt. Despite having a wealth of artworks from this period in collections worldwide, the number of analytical studies on the organic media of these pieces is limited, especially since micro-samples are often required. Over the past two decades, the introduction of high-resolution tandem mass spectrometry (MS)-based proteomics has allowed confident identification of ancient protein remains. In this study, we used MS-based proteomics to analyse micro-samples from multiple ancient Egyptian artefacts, such as painted wooden coffins, cartonnages, wall paintings, and limestone architectural elements, dating 1425 BCE – 400 CE, from the collections of the Medelhavsmuseet in Stockholm (Sweden), the British Museum in London (UK) and the Ny Carlsberg Glyptotek in Copenhagen (Denmark). Our analytical approach allowed the identification of traditional proteinaceous binders and adhesives, such as egg and glue, as well as their species of origin. Interestingly, MS-based proteomics also revealed the unexpected presence of seed storage proteins from sesame and moringa and other plant proteins previously undocumented in artworks from ancient Egypt. Their identification might be the first taxonomical confirmation of the presence and use of sesame and moringa products in ancient Egypt. This prompts new ideas about the origin of the materials, their intentionality and connection with deities, and their interpretation within the artistic, cultural and economic contexts of ancient Egypt. The findings in this research demonstrate how proteomics can provide new evidence to advance the understanding of the biological origin and use of protein sources in art, along with identifying unconventional materials that could not have been detected with other analytical strategies. Our study demonstrates that combining scientific data with information from archeological evidence and textual sources, is necessary for understanding the nature of binding media and adhesives in ancient Egypt.