Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, ON, Canada.
Clin Biochem. 2012 Feb;45(3):223-30. doi: 10.1016/j.clinbiochem.2011.11.015. Epub 2011 Dec 14.
Biomarkers are urgently needed for diagnosis, prognosis and monitoring of lung transplant chronic graft dysfunction. Bronchoalveolar lavage fluid (BAL) has been used in the past as proximal fluid for biomarker discovery in various lung diseases including chronic graft dysfunction (CGD). The current study describes the proteomic analysis of BAL fluids collected from 4 asymptomatic post-transplant patients and 3 patients with symptoms of CGD.
BAL proteome was fractionated by size-exclusion chromatography at protein level and reverse-phase-chromatography at peptide level followed by Orbitrap mass spectrometry detection.
Our in-depth proteomic analysis identified 531 proteins, the largest catalog of BAL proteins reported to date in the context of CGD. A total of 30 and 39 proteins detected exclusively in CGD and non-CGD samples, respectively, are potential candidates for verification phase.
A new protocol was developed to enhance the sensitivity of detecting less abundant proteins in BAL.
生物标志物对于肺移植慢性移植物功能障碍的诊断、预后和监测至关重要。过去，支气管肺泡灌洗液（BAL）已被用于各种肺部疾病（包括慢性移植物功能障碍[CGD]）的生物标志物发现的近端液体。本研究描述了从 4 名无症状移植后患者和 3 名有 CGD 症状患者的 BAL 液中收集的蛋白质组学分析。
BAL 蛋白质组在蛋白质水平上通过分子筛色谱法和肽水平上的反相色谱法进行分级，然后进行 Orbitrap 质谱检测。
我们的深入蛋白质组学分析鉴定了 531 种蛋白质，这是迄今为止在 CGD 背景下报告的 BAL 蛋白质的最大目录。在 CGD 和非 CGD 样本中分别检测到 30 种和 39 种独特的蛋白质，它们是验证阶段的潜在候选物。
开发了一种新方案来提高 BAL 中检测较少丰度蛋白质的灵敏度。
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