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    • 22. 发明申请
    • Multiplex MRM Assay for Evaluation of Cancer
    • 用于评估癌症的多重MRM测定
    • US20160274124A1
    • 2016-09-22
    • US15174970
    • 2016-06-06
    • Expression Pathology, Inc.
    • David B. KRIZMANTodd HEMBROUGHSheeno THYPARAMBILWei-Li LIAO
    • G01N33/68
    • G01N33/6848G01N33/6893G01N33/743G01N2333/47G01N2333/70567G01N2800/56G01N2800/7028
    • The current disclosure provides specific peptides, and derived ionization characteristics of the peptides from the estrogen receptor (ER), progesterone receptor (PR), and/or antigen Ki67 (Ki67) proteins that are particularly advantageous for quantifying the ER, PR, and/or Ki67 proteins directly in biological samples that have been fixed in formalin by the method of Selected Reaction Monitoring/Multiple Reaction Monitoring (SRM/MRM) mass spectrometry. Such biological samples are chemically preserved and fixed wherein the biological sample is selected from tissues and cells treated with formaldehyde containing agents/fixatives including formalin-fixed tissue/cells, formalin-fixed/paraffin embedded (FFPE) tissue/cells, FFPE tissue blocks and cells from those blocks, and tissue culture cells that have been formalin fixed and or paraffin embedded. A protein sample is prepared from a biological sample using the Liquid Tissue™ reagents and protocol, and the ER, PR, and/or Ki67 proteins are quantitated in the Liquid Tissue™ sample by the method of SRM/MRM mass spectrometry by quantitating in the protein sample at least one or more of the peptides described for one or more of the ER, PR, and/or Ki67 proteins. These peptides can be quantitated if they reside in a modified or in an unmodified form. An example of a modified form of an ER, PR, and/or Ki67 peptide is phosphorylation of a tyrosine, threonine, serine, and/or other amino acid residues within the peptide sequence.
    • 目前的公开内容提供特异性肽,以及来自雌激素受体(ER),孕酮受体(PR)和/或抗原Ki67(Ki67)蛋白质的肽的衍生电离特征,其特别有利于定量ER,PR和/ 或通过选择反应监测/多反应监测(SRM / MRM)质谱法的方法在已经在福尔马林中固定的生物样品中直接存在Ki67蛋白。 这些生物样品被化学保存和固定,其中生物样品选自用含甲醛的试剂/固定剂(包括福尔马林固定的组织/细胞,福尔马林固定/石蜡包埋的(FFPE)组织/细胞,FFPE组织块)和 来自这些区块的细胞和已被福尔马林固定和石蜡包埋的组织培养细胞。 使用Liquid Tissue TM试剂和方案从生物样品制备蛋白质样品,并通过SRM / MRM质谱法的方法在液体组织样品中定量分析ER,PR和/或Ki67蛋白质, 蛋白质样品至少一种或多种针对一种或多种ER,PR和/或Ki67蛋白质描述的肽。 如果它们以修饰或未修饰的形式存在,则可以定量这些肽。 ER,PR和/或Ki67肽的修饰形式的实例是肽序列内的酪氨酸,苏氨酸,丝氨酸和/或其他氨基酸残基的磷酸化。