Proteome analysis of microdissected formalin-fixed and paraffin-embedded tissue specimens
Proteome analysis of microdissected formalin-fixed and paraffin-embedded tissue specimens
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DOI：
10.1369/jhc.7a7177.2007
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发表时间：
2007-07-01
影响因子：
3.2
通讯作者：
Balgley, Brian M.
中科院分区：
文献类型：
--
作者：
Guo, Tong;Wang, Weijie;Balgley, Brian M.
靶向蛋白质组学研究基于从高质量组织样本中选择的分离细胞群中丰富与疾病相关的蛋白质，为识别用于临床环境以及在临床前测试和临床试验期间使用的诊断、预后和预测性生物标志物以及发现和验证新的蛋白质药物靶点提供了巨大的潜力。福尔马林固定和石蜡包埋(FFPE)组织收集，以及附加的临床和结果信息，是进行回溯性蛋白质生物标记物研究和进行癌症和其他疾病的翻译研究的宝贵资源。采用毛细管等电聚焦/纳米反相液相色谱与纳米电喷雾电离-串联质谱仪相结合的方法，研究了热诱导抗原修复(AR)技术从显微切割的FFPE胶质母细胞瘤组织中提取的蛋白质。总共识别了14,478个不同的多肽，导致识别出2733个非冗余的SwissProt蛋白质条目。鉴定出的83%的FFPE组织蛋白与从同一患者新鲜冰冻组织的颗粒部分中获得的蛋白重叠。这种高度的蛋白质重叠归因于基于洗涤剂的蛋白质提取在细胞颗粒制备方案和AR技术中的应用。
Targeted proteomics research, based on the enrichment of disease-relevant proteins from isolated cell populations selected from high-quality tissue specimens, offers great potential for the identification of diagnostic, prognostic, and predictive biological markers for use in the clinical setting and during preclinical testing and clinical trials, as well as for the discovery and validation of new protein drug targets. Formalin-fixed and paraffin embedded (FFPE) tissue collections, with attached clinical and outcome information, are invaluable resources for conducting retrospective protein biomarker investigations and performing translational studies of cancer and other diseases. Combined capillary isoelectric focusing/nano-reversed-phase liquid chromatography separations equipped with nano-electrospray ionization-tandem mass spectrometry are employed for the studies of proteins extracted from microdissected FFPE glioblastoma tissues using a heat-induced antigen retrieval (AR) technique. A total of 14,478 distinct peptides are identified, leading to the identification of 2733 non-redundant SwissProt protein entries. Eighty-three percent of identified FFPE tissue proteins overlap with those obtained from the pellet fraction of fresh-frozen tissue of the same patient. This large degree of protein overlapping is attributed to the application of detergent-based protein extraction in both the cell pellet preparation protocol and the AR technique.