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细胞微阵列中蛋白质表达分析:基于抗体的蛋白质组学工具。

Analysis of protein expression in cell microarrays: a tool for antibody-based proteomics.

作者信息

Andersson Ann-Catrin, Strömberg Sara, Bäckvall Helena, Kampf Caroline, Uhlen Mathias, Wester Kenneth, Pontén Fredrik

机构信息

Department of Genetics and Pathology, Rudbeck Laboratory, University Hospital, Uppsala, Sweden.

出版信息

J Histochem Cytochem. 2006 Dec;54(12):1413-23. doi: 10.1369/jhc.6A7001.2006. Epub 2006 Sep 6.

Abstract

Tissue microarray (TMA) technology provides a possibility to explore protein expression patterns in a multitude of normal and disease tissues in a high-throughput setting. Although TMAs have been used for analysis of tissue samples, robust methods for studying in vitro cultured cell lines and cell aspirates in a TMA format have been lacking. We have adopted a technique to homogeneously distribute cells in an agarose gel matrix, creating an artificial tissue. This enables simultaneous profiling of protein expression in suspension- and adherent-grown cell samples assembled in a microarray. In addition, the present study provides an optimized strategy for the basic laboratory steps to efficiently produce TMAs. Presented modifications resulted in an improved quality of specimens and a higher section yield compared with standard TMA production protocols. Sections from the generated cell TMAs were tested for immunohistochemical staining properties using 20 well-characterized antibodies. Comparison of immunoreactivity in cultured dispersed cells and corresponding cells in tissue samples showed congruent results for all tested antibodies. We conclude that a modified TMA technique, including cell samples, provides a valuable tool for high-throughput analysis of protein expression, and that this technique can be used for global approaches to explore the human proteome.

摘要

组织微阵列(TMA)技术为在高通量条件下探索多种正常和疾病组织中的蛋白质表达模式提供了可能。尽管TMA已用于组织样本分析,但一直缺乏以TMA形式研究体外培养细胞系和细胞吸出物的可靠方法。我们采用了一种技术,使细胞在琼脂糖凝胶基质中均匀分布,从而创建人工组织。这使得能够同时分析以微阵列形式组装的悬浮生长和贴壁生长细胞样本中的蛋白质表达。此外,本研究为有效生产TMA的基础实验室步骤提供了优化策略。与标准TMA生产方案相比,所提出的改进措施提高了标本质量并增加了切片产量。使用20种特征明确的抗体对所生成的细胞TMA切片进行免疫组织化学染色特性测试。对培养的分散细胞和组织样本中相应细胞的免疫反应性比较显示,所有测试抗体的结果均一致。我们得出结论,包括细胞样本在内的改良TMA技术为蛋白质表达的高通量分析提供了有价值的工具,并且该技术可用于探索人类蛋白质组的整体方法。

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