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SRM/MRM Assay for the Tumor Necrosis Factor Receptor Superfamily Member 8 (CD30) Protein

机译:肿瘤坏死因子受体超家族成员8(CD30)蛋白的SRM / MRM分析

摘要

The current disclosure provides for specific peptides, and derived ionization characteristics of the peptides, from the tumor necrosis factor receptor superfamily member 8 protein (CD30) that are particularly advantageous for quantifying the CD30 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 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 the biological sample using the Liquid Tissue reagents and protocol and the CD30 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 a CD30 peptide is phosphorylation of a tyrosine, threonine, serine, and/or other amino acid residues within the peptide sequence.
机译:本公开提供了来自肿瘤坏死因子受体超家族成员8蛋白(CD30)的特定肽和衍生的肽的电离特性,其特别有利于直接定量已经被福尔马林固定在福尔马林中的生物样品中的CD30蛋白。选择性反应监测(SRM)质谱法,也可以称为多反应监测(MRM)质谱法。此类生物样品经过化学保存和固定,其中生物样品选自用含甲醛的试剂/固定剂处理过的组织和细胞,包括甲醛固定的组织/细胞,甲醛固定/石蜡包埋的(FFPE)组织/细胞,FFPE组织块和那些块中的细胞,以及已被福尔马林固定和/或石蜡包埋的组织培养细胞。使用液体组织试剂和规程从生物样品中制备蛋白质样品,并通过SRM / MRM质谱法通过在蛋白质样品中定量至少一种或多种肽,在液体组织样品中对CD30蛋白质进行定量描述。如果这些肽以修饰或未修饰的形式存在,则可以定量。 CD30肽的修饰形式的一个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。

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