Elucidation of cellular processes and their changes at the level of protein expression and post-translational modification patterns may allow identification of novel proteins and thereby mechanisms involved in the pathogenesis of multigenic diseases. The aim of this study was to test cultured, nontransformed primary fibroblasts derived from human skin biopsies as a suitable model system for proteome analysis. Therefore soluble protein fractions were separated on several overlapping ultrazoom gels covering the pH range from 3.5-9. Correlation analysis of gel-pairs revealed a highly reproducible protein expression pattern within (intra-assay) and between (inter-assay) independent experiments of a single fibroblast cell line (intra-cell line comparison). Spot intensity variations were less than a factor of two for more than 80% of identical spots. In addition, inter-cell line comparison exhibits no significant variations in spot intensities. To achieve further improvements in reproducibility we generated master gels for each pH range by combining averaged spot information derived from two different cell lines each analysed by two independent experiments using the raw master gel algorithm of the Z3 image analysis software. The resulting reference images of primary human fibroblasts provided a basis for investigating regulation by extracellular stimuli and drugs as well as their alterations in patients with different diseases.

中文翻译：

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初级皮肤成纤维细胞作为蛋白质组分析的人类模型系统。

阐明细胞过程及其在蛋白质表达水平和翻译后修饰模式上的变化可能允许鉴定新蛋白质，从而鉴定出与多基因疾病发病机理有关的机制。这项研究的目的是测试源自人类皮肤活检组织的培养的，未转化的原代成纤维细胞，作为蛋白质组学分析的合适模型系统。因此，在覆盖pH范围为3.5-9的几种重叠的超变焦凝胶上分离了可溶性蛋白质级分。凝胶对的相关性分析揭示了单个成纤维细胞系在（测定内）和（测定间）独立实验之间（测定内）的蛋白表达模式具有高度可重复性。对于超过80％的相同斑点，斑点强度变化小于两倍。另外，小区间细胞系比较显示斑点强度没有显着变化。为了实现可重复性的进一步改善，我们通过组合源自两个不同细胞系的平均斑点信息（使用Z3图像分析软件的原始主胶算法通过两次独立实验进行分析）来生成每个pH范围的主胶。由此产生的人类原代成纤维细胞参考图像为研究细胞外刺激和药物的调控及其在不同疾病患者中的变化提供了基础。为了实现可重复性的进一步改善，我们通过组合源自两个不同细胞系的平均斑点信息（使用Z3图像分析软件的原始主胶算法通过两次独立实验进行分析）来生成每个pH范围的主胶。由此产生的人类原代成纤维细胞参考图像为研究细胞外刺激和药物及其在不同疾病患者中的改变提供了基础。为了实现可重复性的进一步改善，我们通过组合源自两个不同细胞系的平均斑点信息（使用Z3图像分析软件的原始主胶算法通过两次独立实验进行分析）来生成每个pH范围的主胶。由此产生的人类原代成纤维细胞参考图像为研究细胞外刺激和药物的调控及其在不同疾病患者中的变化提供了基础。