Due to methodological advances in the archaeological sciences, an increasing number of archaeological specimens undergo destructive sampling. However, the preservation of cultural heritage is a primary concern. This leads to a dilemma between accessing sample material and obtaining sufficient information for a meaningful analytical outcome. Ideally, sampling a specimen would preserve the object for further macro, micro, and molecular analyses. For palaeoproteomics, a number of minimally invasive sampling approaches have been proposed, representing different benefits and limitations. There have been studies comparing a selection of these protocols, however, these have focused on specimens from a homogenous preservation environment using Zooarchaeology by Mass Spectrometry (ZooMS). Here we expand on earlier work by extending the comparison to specimens from two highly different preservation environments through both ZooMS and liquid-chromatography tandem mass spectrometry (LC-MS/MS). We compare five sampling approaches and seven extraction protocols in total, on 10 sp. humeri from the Early Neolithic site of La Draga, Spain, utilising MALDI-ToF MS and LC-MS/MS to generate proteomic output, while assessing protocol invasiveness using microscopy and 3D imaging. Five humeri originate from Sector A, which is mostly related to dry, terrestrial preservation conditions, while the other five humeri stem from Sector B, which is characterised by its phreatic/aquatic preservation conditions. We show that there is a significant difference in protein recovery and taxonomic specificity between the sampling techniques applied, as well as between burial conditions. Additionally, various surface modifications were observed depending on the specific sampling technique applied. It is therefore essential to assess protein preservation for each sedimentological context within an archaeological site before performing extensive sampling, as protein preservation can be highly inter- and intra-site-specific.

中文翻译：

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通过古蛋白质组学分析新石器时代早期肱骨的市售微创采样方法的比较研究


由于考古科学方法的进步，越来越多的考古标本接受破坏性采样。然而，文化遗产的保护是首要问题。这导致在获取样本材料和获取足够的信息以获得有意义的分析结果之间陷入困境。理想情况下，对样本进行采样可以保存物体以进行进一步的宏观、微观和分子分析。对于古蛋白质组学，已经提出了许多微创采样方法，代表了不同的优点和局限性。已经有研究比较了这些协议的选择，然而，这些研究重点是使用质谱动物考古学 (ZooMS) 来自同质保存环境的标本。在这里，我们通过 ZoomMS 和液相色谱串联质谱 (LC-MS/MS) 扩展了对来自两个高度不同保存环境的标本的比较，从而扩展了早期工作。我们在 10 个 sp 上总共比较了五种采样方法和七种提取方案。西班牙拉德拉加新石器时代早期遗址的肱骨，利用 MALDI-ToF MS 和 LC-MS/MS 生成蛋白质组输出，同时使用显微镜和 3D 成像评估方案侵入性。五个肱骨来自 A 区，主要与干燥、陆地保存条件有关，而其他五个肱骨来自 B 区，其特点是潜水/水生保存条件。我们表明，所应用的采样技术之间以及埋葬条件之间的蛋白质回收率和分类特异性存在显着差异。此外，根据所采用的具体采样技术，观察到了各种表面修饰。 因此，在进行广泛采样之前，有必要评估考古遗址内每种沉积环境的蛋白质保存情况，因为蛋白质保存可能具有高度的遗址间和遗址内特异性。