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Methods and devices for the isolation and analysis of cellular protein content

机译:分离和分析细胞蛋白含量的方法和装置

摘要

The present invention describes devices and methods for performing protein analysis on laser capture microdissected cells, which permits proteomic analysis on cells of different populations. Particular disclosed examples are analysis of normal versus malignant cells, or a comparison of differential protein expression in cells that are progressing from normal to malignant. The protein content of the microdissected cells may be analyzed using techniques such as immunoassays, 1D and 2D gel electrophoresis characterization, Western blotting, liquid chromatography quadrapole ion trap electrospray (LCQ-MS), Matrix Assisted Laser Desorption Ionization/Time of Flight (MALDI/TOF), and Surface Enhanced Laser Desorption Ionization Spectroscopy (SELDI). In addition to permitting direct comparison of qualitative and quantitative protein content of tumor cells and normal cells from the same tissue sample, the methods also allow for investigation of protein characteristics of tumor cells, such as binding ability and amino acid sequence, and differential expression of proteins in particular cell populations in response to drug treatment. The present methods also provide, through the use of protein fingerprinting, a rapid and reliable way to identify the source tissue of a tumor metastasis.
机译:本发明描述了用于在激光捕获显微切割的细胞上进行蛋白质分析的设备和方法,其允许对不同群体的细胞进行蛋白质组学分析。具体公开的例子是正常细胞与恶性细胞的分析,或从正常细胞发展到恶性细胞的差异蛋白质表达的比较。可以使用免疫分析,一维和二维凝胶电泳鉴定,蛋白质印迹,液相色谱四极离子阱电喷雾(LCQ-MS),基质辅助激光解吸电离/飞行时间(MALDI / TOF)和表面增强激光解吸电离光谱(SELDI)。除了可以直接比较来自同一组织样本的肿瘤细胞和正常细胞的定性和定量蛋白质含量外,这些方法还可以研究肿瘤细胞的蛋白质特征,例如结合能力和氨基酸序列,以及肿瘤细胞的差异表达。药物处理后特定细胞群体中的蛋白质。本发明的方法还通过使用蛋白质指纹图谱提供了一种快速可靠的方法来鉴定肿瘤转移的来源组织。

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