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*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

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.

Original languageEnglish
Article number108191
JournaliScience
Volume26
Issue number11
DOIs
Publication statusPublished - 17 Nov 2023

Bibliographical note

Funding Information:
This work was supported by the University of Nottingham Interdisciplinary Center for Analytical Science ( UNICAS ). The Arnhem skeletal collection is generously on loan to the Laboratory for Human Osteoarchaeology at Leiden University from Gemeente Arnhem and we thank Martijn Defilet for access to the collection. The archaeological excavation at Tiel Medel was executed by a combination of archaeological firms: RAAP (Weesp), ADC ArcheoProjecten (Amersfoort), and Archol (Leiden). Osteoarchaeological research on the inhumations was carried out by Steffen Baetsen. Theo ten Anscher (RAAP) is thanked for providing context information. Excavation of the St Peter’s Churchyard Cemetery were directed by Headland Archaeology (Edinburgh), who gave permission for the St Peter’s remains to be included in this study. The authors thank University of Nottingham project students that contributed to early method development: Elspeth Bell; Dylan Nicol; Luke Boy; Hannah Fane; Ellen Wikeley; George Parrington; Daniel Youngs; Lina Asobirionwu; Richard Svitan; Sandra Frost; Charlotte Rayner. This work is dedicated to the memory of Andy Barlow (Edinburgh).

Funding Information:
This work was supported by the University of Nottingham Interdisciplinary Center for Analytical Science (UNICAS). The Arnhem skeletal collection is generously on loan to the Laboratory for Human Osteoarchaeology at Leiden University from Gemeente Arnhem and we thank Martijn Defilet for access to the collection. The archaeological excavation at Tiel Medel was executed by a combination of archaeological firms: RAAP (Weesp), ADC ArcheoProjecten (Amersfoort), and Archol (Leiden). Osteoarchaeological research on the inhumations was carried out by Steffen Baetsen. Theo ten Anscher (RAAP) is thanked for providing context information. Excavation of the St Peter's Churchyard Cemetery were directed by Headland Archaeology (Edinburgh), who gave permission for the St Peter's remains to be included in this study. The authors thank University of Nottingham project students that contributed to early method development: Elspeth Bell; Dylan Nicol; Luke Boy; Hannah Fane; Ellen Wikeley; George Parrington; Daniel Youngs; Lina Asobirionwu; Richard Svitan; Sandra Frost; Charlotte Rayner. This work is dedicated to the memory of Andy Barlow (Edinburgh). R.L. and S.S. developed the overall concept of the study; B.S. P.H.S. C.P. L.F. P.T. L.M.K, S.S. and R.L. designed research; B.S. S.F. P.H.S. A.B. N.O. and P.T. performed research; C.P. L.F. L.M.K, S.S. P.H.S. and R.L. analyzed data; P.H.S. C.P. L.F. N.O. P.T. L.M.K. S.S. and R.L. wrote the paper; P.H.S. and R.L. prepared the data presentation. The authors declare no competing interests.

Publisher Copyright:
© 2023 The Author(s)

Funding

This work was supported by the University of Nottingham Interdisciplinary Center for Analytical Science ( UNICAS ). The Arnhem skeletal collection is generously on loan to the Laboratory for Human Osteoarchaeology at Leiden University from Gemeente Arnhem and we thank Martijn Defilet for access to the collection. The archaeological excavation at Tiel Medel was executed by a combination of archaeological firms: RAAP (Weesp), ADC ArcheoProjecten (Amersfoort), and Archol (Leiden). Osteoarchaeological research on the inhumations was carried out by Steffen Baetsen. Theo ten Anscher (RAAP) is thanked for providing context information. Excavation of the St Peter’s Churchyard Cemetery were directed by Headland Archaeology (Edinburgh), who gave permission for the St Peter’s remains to be included in this study. The authors thank University of Nottingham project students that contributed to early method development: Elspeth Bell; Dylan Nicol; Luke Boy; Hannah Fane; Ellen Wikeley; George Parrington; Daniel Youngs; Lina Asobirionwu; Richard Svitan; Sandra Frost; Charlotte Rayner. This work is dedicated to the memory of Andy Barlow (Edinburgh). This work was supported by the University of Nottingham Interdisciplinary Center for Analytical Science (UNICAS). The Arnhem skeletal collection is generously on loan to the Laboratory for Human Osteoarchaeology at Leiden University from Gemeente Arnhem and we thank Martijn Defilet for access to the collection. The archaeological excavation at Tiel Medel was executed by a combination of archaeological firms: RAAP (Weesp), ADC ArcheoProjecten (Amersfoort), and Archol (Leiden). Osteoarchaeological research on the inhumations was carried out by Steffen Baetsen. Theo ten Anscher (RAAP) is thanked for providing context information. Excavation of the St Peter's Churchyard Cemetery were directed by Headland Archaeology (Edinburgh), who gave permission for the St Peter's remains to be included in this study. The authors thank University of Nottingham project students that contributed to early method development: Elspeth Bell; Dylan Nicol; Luke Boy; Hannah Fane; Ellen Wikeley; George Parrington; Daniel Youngs; Lina Asobirionwu; Richard Svitan; Sandra Frost; Charlotte Rayner. This work is dedicated to the memory of Andy Barlow (Edinburgh). R.L. and S.S. developed the overall concept of the study; B.S. P.H.S. C.P. L.F. P.T. L.M.K, S.S. and R.L. designed research; B.S. S.F. P.H.S. A.B. N.O. and P.T. performed research; C.P. L.F. L.M.K, S.S. P.H.S. and R.L. analyzed data; P.H.S. C.P. L.F. N.O. P.T. L.M.K. S.S. and R.L. wrote the paper; P.H.S. and R.L. prepared the data presentation. The authors declare no competing interests.

Keywords

  • Immunology
  • Methodology in biological sciences
  • Paleobiology

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