Of Patients Like No Other: High‐Resolution Paleoproteomic Analysis of Biological Residues on Garments Attributed to Marie‐Antoinette and Louis XVII

RATIONALE: Paleoproteomic analysis of biological traces present on the clothing of Queen Marie-Antoinette » (1755 -1793) and Louis Charles de France, Duke of Normandie and future Louis XVII (1785-1795) has made it possible to precisely identify the composition of these residues and to better understand the health status of these two "patients from the past." METHODS: Proteins were extracted from 16 samples collected from six different clothing items, proteolyzed with trypsin, and the resulting peptides were analyzed by tandem mass spectrometry using an Orbitrap Astral instrument. An initial proteotyping analysis, performed without any prior assumptions regarding species origin, unambiguously confirmed the proteins to be of human origin. A subsequent targeted database search was then conducted for protein identification and label-free quantification. RESULTS: A total of 76 421 peptide-to-spectrum matches were obtained, and 3457 proteotypic peptides were listed, allowing the identification of 361 proteins with at least two specific peptide sequences per protein in each individual sample. PAUF and STIP1 peptides were detected in residues attributed to Marie-Antoinette, whereas proteins associated with male urogenital tissues and inflammatory responses were detected in residues attributed to Louis XVII. These findings are compatible with blood- or urine-containing residues and nonspecific neoplastic or inflammatory contexts, respectively. CONCLUSION: This study demonstrates the potential of paleoproteomics for reconstructing paleopathological profiles from biological traces on ancient materials, opening new perspectives in the study of disease history, paleoepidemiology, and retrospective diagnosis.