Anal Chem. 2010 Mar 1;82(5):1584-8. doi: 10.1021/ac902204k.
A method that relies on subtractive tissue-directed shot-gun proteomics to identify tumor proteins in the blood of a patient newly diagnosed with cancer is described. To avoid analytical and statistical biases caused by physiologic variability of protein expression in the human population, this method was applied on clinical specimens obtained from a single patient diagnosed with nonmetastatic renal cell carcinoma (RCC). The proteomes extracted from tumor, normal adjacent tissue and preoperative plasma were analyzed using 2D-liquid chromatography-mass spectrometry (LC-MS). The lists of identified proteins were filtered to discover proteins that (i) were found in the tumor but not normal tissue, (ii) were identified in matching plasma, and (iii) whose spectral count was higher in tumor tissue than plasma. These filtering criteria resulted in identification of eight tumor proteins in the blood. Subsequent Western-blot analysis confirmed the presence of cadherin-5, cadherin-11, DEAD-box protein-23, and pyruvate kinase in the blood of the patient in the study as well as in the blood of four other patients diagnosed with RCC. These results demonstrate the utility of a combined blood/tissue analysis strategy that permits the detection of tumor proteins in the blood of a patient diagnosed with RCC.
一种依赖于组织导向的消减 shotgun 蛋白质组学的方法，用于鉴定新诊断为癌症的患者血液中的肿瘤蛋白。为了避免由于人群中蛋白质表达的生理变异性引起的分析和统计偏差，该方法应用于从单个诊断为非转移性肾细胞癌（RCC）的患者获得的临床标本。使用二维液相色谱-质谱（LC-MS）分析从肿瘤、正常相邻组织和术前血浆中提取的蛋白质组。鉴定的蛋白质列表经过过滤，以发现（i）仅在肿瘤中而不在正常组织中发现的蛋白质，（ii）在匹配的血浆中鉴定的蛋白质，以及（iii）在肿瘤组织中的光谱计数高于血浆中的蛋白质。这些过滤标准导致在研究中的患者的血液中鉴定出 8 种肿瘤蛋白。随后的 Western-blot 分析证实了研究中患者以及另外 4 名诊断为 RCC 的患者的血液中存在钙粘蛋白-5、钙粘蛋白-11、DEAD 盒蛋白-23 和丙酮酸激酶。这些结果证明了联合血液/组织分析策略的有效性，该策略允许检测诊断为 RCC 的患者血液中的肿瘤蛋白。
