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针对肺癌生物标志物发现的人支气管肺泡灌洗液深度蛋白质组学分析

In-Depth Proteomic Analysis of Human Bronchoalveolar Lavage Fluid toward the Biomarker Discovery for Lung Cancers.

作者信息

Sim Seo Young, Choi Yu Ri, Lee Jun Hyung, Lim Jae Min, Lee Seung-Eun, Kim Kwang Pyo, Kim Jin Young, Lee Seung Hyeun, Kim Min-Sik

机构信息

Department of Applied Chemistry, Global Center for Pharmaceutical Ingredient Materials, Institute of Natural Science, Kyung Hee University, Yongin, 17104, Republic of Korea.

Department of New Biology, DGIST, Daegu, 42988, Republic of Korea.

出版信息

Proteomics Clin Appl. 2019 Sep;13(5):e1900028. doi: 10.1002/prca.201900028. Epub 2019 Jun 6.

Abstract

PURPOSE

Lung cancer is among the most common cancers. Bronchoalveolar lavage fluid (BALF) can be easily obtained from patients with lung cancers. The aim is to develop a novel proteomic method of the molecule-based sensitive detection of biomarkers from BALF.

EXPERIMENTAL DESIGN

BALF samples are collected from segmental bronchus of 14 patients with lung cancers from Kyung Hee University Hospital. First, BALF proteome is depleted using a depletion column, and then peptides are prepared from the enriched low abundant proteins and fractionated by high pH reverse phase liquid chromatography prior to LC-MS/MS. Data are available via ProteomeXchange with identifier PXD012645.

RESULTS

A novel method is developed for in-depth proteomic analysis of BALF by combining antibody-based depletion of high abundant proteins from BALF with high pH peptide fractionation. Peptides are analyzed on a high resolution Orbitrap Fusion mass spectrometer. MaxQuant search result in the identification of 4615 protein groups mapped to 4534 genes.

CONCLUSIONS AND CLINICAL RELEVANCE

It is found that the method outperformed conventional BALF proteomic methods and it is believed that this method will facilitate the biomarker research for lung cancer. In addition, it is shown that BALF will be a great source of biomarkers of lung diseases.

摘要

目的

肺癌是最常见的癌症之一。支气管肺泡灌洗液(BALF)可从肺癌患者中轻松获取。目的是开发一种基于分子的新型蛋白质组学方法,用于从BALF中灵敏检测生物标志物。

实验设计

从庆熙大学医院的14例肺癌患者的节段支气管收集BALF样本。首先,使用去除柱去除BALF蛋白质组,然后从富集的低丰度蛋白质中制备肽,并在进行液相色谱-串联质谱分析之前通过高pH反相液相色谱进行分离。数据可通过ProteomeXchange获取,标识符为PXD012645。

结果

通过将基于抗体的从BALF中去除高丰度蛋白质与高pH肽分离相结合,开发了一种用于BALF深度蛋白质组学分析的新方法。在高分辨率的Orbitrap Fusion质谱仪上分析肽。MaxQuant搜索结果鉴定出4615个蛋白质组,对应4534个基因。

结论与临床意义

发现该方法优于传统的BALF蛋白质组学方法,并且认为该方法将促进肺癌生物标志物的研究。此外,结果表明BALF将是肺部疾病生物标志物的重要来源。

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