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Tiny Hopes─Assessing Protein Preservation in Collagen-Depleted Bones
Emese I Végh1,2, Leila Afjehi-Sadat3, Maddalena Giannì1,2
1Department of Evolutionary Anthropology, University of Vienna, 1 Djerassiplatz, A-1030 Vienna, Austria.
Journal of Proteome Research
|June 12, 2026
Summary
Ancient bones, even those appearing collagen-free, often retain measurable collagen. This finding expands biomolecular analysis possibilities for human evolution studies in warm climates.
Area of Science:
- Paleoproteomics
- Archaeological Science
- Human Evolution
Background:
- Biomolecular data from ancient skeletal tissues is crucial for understanding human evolution.
- Warm climates often lead to complete collagen loss in bones, limiting study regions.
- Traditional methods may misclassify bones as collagen-free, excluding them from valuable analyses.
Purpose of the Study:
- To investigate the survival of non-collagenous proteins (NCPs) and low-molecular-weight proteins in archaeological bones.
- To assess protein preservation in bones traditionally deemed collagen-free.
- To refine methods for protein extraction and analysis from challenging ancient samples.
Main Methods:
- Sandwich enzyme-linked immunosorbent assays (ELISAs) for osteocalcin quantification.
- Qubit fluorometry for total protein measurement.
- Liquid chromatography tandem mass spectrometry (LC-MS/MS) for protein characterization.
- Fourier transform infrared spectroscopy (FTIR) for diagenetic assessment.
- Comparison of tangential flow filtration and centrifugal devices for protein recovery.
- Evaluation of EDTA and HCl demineralization techniques with FASP and protein precipitation.
Main Results:
- Non-collagenous proteins (NCPs), including osteocalcin, survive only if insoluble collagen is preserved.
- Collagen was detected in the soluble supernatants of most "collagen-depleted" bones.
- Abundant collagen peptides were found even in samples with 0% traditional collagen yield.
- Tangential flow filtration showed superior protein recovery compared to centrifugal devices.
- EDTA demineralization with FASP maximized collagen recovery, while HCl demineralization with protein precipitation best detected unique NCPs.
Conclusions:
- Bones unsuitable for traditional collagen dating may still contain measurable collagen.
- This study broadens the scope of biomolecular analyses possible in ancient bones from warm and humid climates.
- Re-evaluation of "collagen-free" bones using advanced proteomic techniques is recommended for human evolution research.
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