Antibody-based sex determination of human skeletal remains

Barry Shaw; Sophie Foggin; Petter Hamilton-Stanley; Andy Barlow; Catriona Pickard; Linda Fibiger; Neil Oldham; Patrick Tighe; Lisette M Kootker; Sarah Schrader; Rob Layfield

doi:10.1016/j.isci.2023.108191

Antibody-based sex determination of human skeletal remains

iScience. 2023 Oct 12;26(11):108191. doi: 10.1016/j.isci.2023.108191. eCollection 2023 Nov 17.

Authors

Affiliations

¹ School of Life Sciences, University of Nottingham, Nottingham, UK.
² School of History, Classics and Archaeology, University of Edinburgh, Edinburgh, UK.
³ School of Chemistry, University of Nottingham, Nottingham, UK.
⁴ Department of Earth Sciences, Vrije Universiteit Amsterdam, Amsterdam, the Netherlands.
⁵ Faculty of Archaeology, Leiden University, Leiden, the Netherlands.

Abstract

Assignment of biological sex to skeletal remains is critical in the accurate reconstruction of the past. Analysis of sex-chromosome encoded AMELX and AMELY peptides from the enamel protein amelogenin underpins a minimally destructive mass spectrometry (MS) method for sex determination of human remains. However, access to such specialist approaches limits applicability. As a convenient alternative, we generated antibodies that distinguish human AMELX and AMELY. Purified antibodies demonstrated high selectivity and quantitative detection against synthetic peptides by ELISA. Using acid etches of enamel from post-medieval skeletons, antibody determinations corrected osteological uncertainties and matched parallel MS, and for Bronze Age samples where only enamel was preserved, also matched MS analyses. Toward improved throughput, automated stations were applied to analyze 19th-century teeth where sex of individuals was documented, confirming MS can be bypassed. Our immunological tools should underpin development of routine, economical, high-throughput methods for sex determination, potentially even in a field setting.

Keywords: Immunology; Methodology in biological sciences; Paleobiology.