Previous work from our laboratories utilized a novel skin taping method and mass spectrometry-based proteomics to discover clinical biomarkers of skin conditions; these included atopic dermatitis, Staphylococcus aureus colonization, and eczema herpeticum. While suitable for discovery purposes, semi-quantitative proteomics is generally time-consuming and expensive. Furthermore, depending on the method used, discovery-based proteomics can result in high variation and inadequate sensitivity to detect low abundant peptides. Therefore, we strove to develop a rapid, sensitive, and reproducible method to quantitate disease-related proteins from skin tapings. We utilized isotopically-labeled peptides and tandem mass spectrometry to obtain absolute quantitation values on 14 peptides from 7 proteins; these proteins had shown previous importance in skin disease. The method demonstrated good reproducibility, dynamic range, and linearity (R² > 0.993) when n = 3 standards were analyzed across 0.05–2.5 pmol. The method was used to determine if differences exist between skin proteins in a small group of atopic versus non-atopic individuals (n = 12). While only minimal differences were found, peptides were detected in all samples and exhibited good correlation between peptides for 5 of the 7 proteins (R² = 0.71–0.98). This method can be applied to larger cohorts to further establish the relationships of these proteins to skin disease.

我们实验室的先前工作是利用一种新颖的皮肤包扎方法和基于质谱的蛋白质组学技术来发现皮肤状况的临床生物标志物。这些包括特应性皮炎，金黄色葡萄球菌定植和湿疹疱疹。半定量蛋白质组学虽然适合发现，但通常既耗时又昂贵。此外，根据使用的方法，基于发现的蛋白质组学可能会导致变异大，而检测低丰度肽段的灵敏度不足。因此，我们努力开发一种快速，灵敏且可重现的方法，以定量从皮肤包扎中与疾病相关的蛋白质。我们利用同位素标记的肽和串联质谱法从7种蛋白质中获得14种肽的绝对定量值。这些蛋白质在皮肤疾病中显示出先前的重要性。该方法具有良好的重现性，动态范围和线性（R ² > 0.993），当n = 3的标准品在0.05–2.5 pmol范围内进行分析。该方法用于确定一小组异位个体与非异位个体（n = 12）中皮肤蛋白之间是否存在差异。虽然只发现了最小的差异，但在所有样品中都检测到了肽，并且在7种蛋白质中的5种之间显示出良好的肽相关性（R ²  = 0.71-0.98）。该方法可用于更大的人群，以进一步建立这些蛋白质与皮肤疾病的关系。