High-Throughput Targeted Paleoproteomics Sex Estimation on Medieval Great Moravian Individuals Using MALDI-CASI-FTICR Mass Spectrometry.

The estimation of the biological sex of archeological remains is crucial information in bioarcheology and forensic anthropology. In recent years, proteomics based on molecular sexual dimorphism has emerged as a preferred method, particularly because of its minimally invasive approach for extracting amelogenin X and Y proteins from tooth enamel. However, there is an increasing demand to accelerate this process and facilitate the analysis of large archeological assemblages. This study presents a novel high-throughput targeted paleoproteomics method for biological sex estimation using MALDI-CASI-FTICR mass spectrometry. This approach combines the strengths of existing methods, including ultrahigh resolution, significantly reduced processing times, targeted analysis, and scalability to large archeological sample sets. The method was initially validated on modern individuals with known sex and subsequently applied to 130 adult and juvenile individuals from medieval Great Moravia (present-day Czech Republic). Biological sex was successfully estimated for all but one of the individuals. The results not only provide a more efficient biological sex estimation but also help to resolve a few errors in sex assessment previously encountered with osteomorphological and tooth morphometric techniques. The implementation of this method significantly improves the accuracy and efficiency of biological sex estimation, offering a powerful tool for anthropological research.