Undertaking the biological sex assessment of human remains: The applicability of minimally-invasive methods for proteomic sex estimation from enamel peptides

Significance The ability to assign biological sex to human skeletal remains is a fundamental requirement in archaeology, paleoanthropology, and medico-legal sciences. While DNA sequencing can be used, it is expensive, time-consuming, and often fails due to the poor quality of the remaining DNA. An easier, more reliable, and consistently applicable method is needed. We present a method for sex determination of human remains using peptides retrieved from tooth enamel. Amelogenin is an enamel-forming protein encoded for by both chromosomes X and Y, with slight differences in their amino acid sequences. Peptides with these differences were identified by nanoflow liquid chromatography mass spectrometry and found to correctly assign sex to archaeological human remains of various chronological ages, from hundreds to thousands of years old.

With the advent of ancient DNA, as well as other methods such as isotope analysis, destructive sampling of archaeofaunal remains has increased much faster than the effort to collect and curate them. While there has been considerable discussion regarding the ethics of destructive sampling and analysis of human remains, this dialogue has not extended to archaeofaunal material. Here we address this gap and discuss the ethical challenges surrounding destructive sampling of materials from archaeofaunal collections. We suggest ways of mitigating the negative aspects of destructive sampling and present step-by-step guidelines aimed at relevant stakeholders, including scientists, holding institutions and scientific journals. Our suggestions are in most cases easily implemented without significant increases in project costs, but with clear long-term benefits in the preservation and use of zooarchaeological material.