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    • 15. 发明申请
    • CALIBRATED RPMA ASSAY
    • 校准RPMA测定
    • WO2009032319A1
    • 2009-03-12
    • PCT/US2008/010445
    • 2008-09-05
    • PETRICOIN III, Emanuel F.LIOTTA, Lance A.
    • PETRICOIN III, Emanuel F.LIOTTA, Lance A.
    • G01N33/96G01N33/53
    • G01N33/96G01N33/5005
    • This invention relates, e.g., to a set of calibrants for determining the amount in a sample of an analyte (e.g., a protein, such as a protein that has been post-translationally modified), comprising a plurality of calibrants, which contain a range of amounts (e.g., defined amounts and/or serial dilutions) of the analyte, spanning the expected amount of the analyte in the sample. In each of the calibrants, a defined amount of the analyte is present in the same suitable, biological diluent (e.g., a cell or tissue lysate, or a bodily fluid). In one embodiment of the invention, the diluent reflects the same or a similar biological milieu (proteins, lipids, serum proteins, serum matrix proteins, etc.) as that in the sample in which the analyte to be measured is present. In embodiments of the invention, a single calibrant (e.g., a cell lysate) may comprise as many as hundreds of analytes, and can be used for the quantification of those hundreds of analytes in a sample. Methods are described for performing an assay (e.g. RPMA analysis), in which the calibrants of a set of calibrants of the invention are immobilized on each of the surfaces to which samples to be analyzed are immobilized, thereby providing an internal calibration curve for quantifying an RPMA assay.
    • 本发明涉及例如用于确定分析物样品中的量(例如蛋白质,例如经翻译后修饰的蛋白质)的一组校准剂,其包含多个校准物,其包含范围 的分析物的量(例如,定义的量和/或连续稀释度),跨越样品中分析物的预期量。 在每个校准中,一定量的分析物存在于相同的合适的生物稀释剂(例如,细胞或组织裂解液或体液)中。 在本发明的一个实施方案中,稀释剂反映与待测分析物存在的样品中相同或相似的生物环境(蛋白质,脂质,血清蛋白,血清基质蛋白等)。 在本发明的实施方案中,单个校准物(例如,细胞裂解物)可以包含多达数百种分析物,并且可以用于量化样品中数百种分析物。 描述了用于进行测定(例如RPMA分析)的方法,其中本发明的一组校准剂的校准物被固定在待分析样品固定的每个表面上,由此提供内部校准曲线,用于量化 RPMA测定。