An improved quantitative mass spectrometry analysis of tumor specific mutant proteins at high sensitivity.
An improved quantitative mass spectrometry analysis of tumor specific mutant proteins at high sensitivity.
复制标题
改进的高灵敏度肿瘤特异性突变蛋白定量质谱分析。
DOI：
10.1002/pmic.201100611
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发表时间：
2012
期刊：
影响因子：
3.4
通讯作者：
Ashman,Keith
中科院分区：
文献类型：
--
作者：
Ruppen-Cañás,Isabel;López-Casas,PedroP;García,Fernando;Ximénez-Embún,Pilar;Muñoz,Manuel;Morelli,MPia;Real,FranciscoX;Serna,Antonio;Hidalgo,Manuel;Ashman,Keith
New disease specific biomarkers, especially for cancer, are urgently needed to improve individual diagnosis, prognosis, and treatment selection, that is, for personalized medicine. Genetic mutations that affect protein function drive cancer. Therefore, the detection of such mutations represents a source of cancer specific biomarkers. Here we confirm the implementation of the mutant protein specific immuno‐SRM (where SRM is selective reaction monitoring) mass spectrometry method of RAS proteins reported by Wang et al. [Proc. Natl. Acad. Sci. USA2011,108,2444–2449], which exploits an antibody to simultaneously capture the different forms of the target protein and the resolving power and sensitivity of LC‐MS/MS and improve the technique by using a more sensitive mass spectrometer. The mutant form G12D was quantified by SRM on a QTRAP 5500 mass spectrometer and the MIDAS workflow was used to confirm the sequence of the targeted peptides. This assay has been applied to quantify wild type and mutant RAS proteins in patient tumors, xenografted human tissue, and benign human epidermal tumors at high sensitivity. The limit of detection for the target proteins was as low as 12 amol (0.25 pg). It requires low starting amounts of tissue (ca.15 mg) that could be obtained from a needle aspiration biopsy. The described strategy could find application in the clinical arena and be applied to the study of expression of protein variants in disease.
