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    • 5. 发明申请
    • INSULIN RECEPTOR SUBSTRATE 1 (IRS1) PROTEIN SRM/MRM ASSAY
    • 胰岛素受体底物1(IRS1)蛋白SRM / MRM测定
    • WO2011087862A1
    • 2011-07-21
    • PCT/US2010/061909
    • 2010-12-22
    • EXPRESSION PATHOLOGY, INC.KRIZMAN, David B.HEMBROUGH, ToddTHYPARAMBIL, Sheeno
    • KRIZMAN, David B.HEMBROUGH, ToddTHYPARAMBIL, Sheeno
    • G01N31/00A61K38/00
    • G01N33/57496G01N33/574G01N33/6848G01N2333/4703G01N2333/62G01N2333/72G01N2440/14G01N2440/38G01N2560/00G01N2800/52
    • The current disclosure provides for specific peptides, and derived ionization characteristics of the peptides, from the Insulin Receptor Substrate 1 (IRS 1 ) protein that are particularly advantageous for quantifying the IRS 1 protein directly in biological samples that have been fixed in formalin by the method of Selected Reaction Monitoring (SRM) mass spectrometry, or what can also be termed as Multiple Reaction Monitoring (MRM) mass spectrometry. Such biological samples are chemically preserved and fixed wherein said 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 said biological sample using the Liquid Tissue™ reagents and protocol and the IRS 1 protein is 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. These peptides can be quantitated if they reside in a modified or an unmodified form. An example of a modified form of an IRS 1 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了来自胰岛素受体底物1(IRS 1)蛋白质的特异性肽和衍生的离子化特征,其特别有利于通过该方法在福尔马林中固定的生物样品中直接定量IRS 1蛋白 的选择反应监测(SRM)质谱,或什么也可以称为多反应监测(MRM)质谱。 这样的生物样品被化学保存和固定,其中所述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 TissueTM试剂和方案从所述生物样品制备蛋白质样品,并且通过SRM / MRM质谱法的方法在液体组织样品中定量IRS 1蛋白质,通过在蛋白质样品中定量至少一种或多种 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。 IRS 1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/或其它氨基酸残基的磷酸化。