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ANALYTICAL PLATFORM AND METHOD FOR GENERATING PROTEIN EXPRESSION PROFILES OF CELL POPULATIONS

机译:生成细胞群体蛋白质表达谱的分析平台和方法

摘要

The present invention is related to analytical platforms and methods performed therewith for generating qualitative and / or quantitative protein expression profiles, in particular differential protein expression profiles, of cell populations comprising: - generating lysates of one or more populations of cells, the lysates comprising a plurality of proteins expressed by the respective cell populations, - providing an essentially planar solid support, - depositing at discrete sites small quantities of the cell lysates, in diluted or undiluted form directly on said solid support or on an adhesion-promoting layer applied on said solid support, thereby creating one or more one- or two-dimensional arrays of discrete measurement areas on said solid support, - applying a number of binding reagents as specific binding partners for the proteins contained in cell lysates in discrete measurement areas and to be detected and, if adequate, one or more detection reagents on said one or more arrays of measurement areas, the binding reagents and the detection reagents being applied sequentially or in a single addition-step, after binding of the detection reagents to the binding reagents, to the one or more arrays of discrete measurement areas for e.g. global analysis of signalling pathways or screening antibody sets/libraries against protein targets for best specificity, selectivity and affinity, and - measuring and recording optical signals emanating from said one or more arrays of discrete measurement areas in a locally resolved manner, wherein said essentially planar solid support is non-porous and an optionally applied adhesion-promoting layer has a thickness of less than 1 µm.
机译:本发明涉及用于产生细胞群体的定性和/或定量蛋白质表达谱,特别是差异蛋白质表达谱的分析平台和方法,所述分析平台和方法包括:-产生一个或多个细胞群的裂解物,所述裂解物包含由各自的细胞群表达的多种蛋白质-提供基本平坦的固体支持物-将少量稀释或未稀释形式的细胞裂解物以离散或稀释的形式直接沉积在所述固体支持物或施加于所述细胞的粘附促进层上固体支持物,从而在所述固体支持物上创建一个或多个一维或二维离散测量区域阵列,-将多种结合试剂用作离散测量区域中细胞裂解物中所含蛋白质的特异性结合伴侣,并进行检测以及在适当的情况下,在所述一个或多个阵列上的一种或多种检测试剂在f个测量区域中,在将检测试剂与结合试剂结合之后,将结合试剂和检测试剂顺序地或在单个添加步骤中施加到例如一个或多个离散测量区域阵列中。全局分析信号通路或针对蛋白质靶标筛选抗体集/文库以获得最佳特异性,选择性和亲和力,以及-以局部分辨的方式测量和记录从所述一个或多个离散测量区域阵列发出的光学信号,其中所述基本上是平面的固体支持物是无孔的,并且可选地应用的增粘层的厚度小于1 µm。

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