Lung cancer serum biomarker discovery using glycoprotein capture and liquid chromatography mass spectrometry.
Lung cancer serum biomarker discovery using glycoprotein capture and liquid chromatography mass spectrometry.
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使用糖蛋白捕获和液相色谱质谱法发现肺癌血清生物标志物。
DOI：
10.1021/pr100696n
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发表时间：
2010-12-03
影响因子：
4.4
通讯作者：
Bigbee, William L.
中科院分区：
文献类型：
--
作者：
Zeng, Xuemei;Hood, Brian L.;Sun, Mai;Conrads, Thomas P.;Day, Roger S.;Weissfeld, Joel L.;Siegfried, Jill M.;Bigbee, William L.
由于众所周知的癌症中蛋白质糖基化模式的改变以及所得糖蛋白质组复杂性的降低，靶向糖蛋白质组学代表了一种基于外周血的癌症生物标志物发现的有吸引力的方法。在这里，我们报告了其在一组混合非小细胞肺癌 (NSCLC) 病例血清（来自 54 名患者的 9 个腺癌和 6 个鳞状细胞癌池）和匹配对照池中的应用，其中包括 8 个临床对照池，计算机断层扫描检测到结节，但通过活检确定为非恶性，来自 54 名患者，以及来自 106 名无癌症受试者的 8 个匹配的健康对照池。该研究的目标是发现可以改善肺癌早期检测和诊断的生物标志物。首先使用免疫亲和扣除法去除最丰富的血清蛋白；然后对剩余的血清蛋白进行基于酰肼化学的糖蛋白捕获和富集。使用酰肼树脂原位胰蛋白酶消化来释放非糖基化肽。以前，N-连接糖基化肽通过肽-N-糖苷酶 F (PNGase F) 处理释放，随后通过液相色谱 (LC)-串联质谱 (MS/MS) 进行分析。开发了基于 MATLAB® 的内部工具，以促进不同 LC-MS/MS 运行之间的保留时间对齐、母离子 m/z 值和洗脱曲线的确定，以及基于已识别肽的确定参数的质量色谱图的积分。来自 22 种不同蛋白质的总共 38 种糖肽在病例/对照池中的丰度存在显着差异（P<0.01，学生 t 检验），并且它们的丰度导致基于层次聚类的病例池和对照池几乎完全分离。其中三种候选蛋白的差异丰度通过池中应用的市售 ELISA 进行了验证。对于所有选定的糖蛋白，观察到糖肽质量色谱图与 ELISA 测量的蛋白质丰度之间存在强正相关性。
Targeted glycoproteomics represents an attractive approach for conducting peripheral blood based cancer biomarker discovery due to the well-known altered pattern of protein glycosylation in cancer and the reduced complexity of the resultant glycoproteome. Here we report its application to a set of pooled non-small cell lung cancer (NSCLC) case sera (9 adenocarcinoma and 6 squamous cell carcinoma pools from 54 patients) and matched controls pools, including 8 clinical control pools with computed tomography detected nodules but being non-malignant as determined by biopsy from 54 patients, and 8 matched healthy control pools from 106 cancer-free subjects. The goal of the study is to discover biomarkers which may enable improved early detection and diagnosis of lung cancer. Immunoaffinity subtraction was used to first deplete the top most abundant serum proteins; the remaining serum proteins were then subjected to hydrazide chemistry based glycoprotein capture and enrichment. Hydrazide resin in situ trypsin digestion was used to release non-glycosylated peptides. Formerly N-linked glycosylated peptides were released by peptide-N-glycosidase F (PNGase F) treatment and were subsequently analyzed by liquid chromatography (LC)-tandem mass spectrometry (MS/MS). A MATLAB® based in-house tool was developed to facilitate retention time alignment across different LC-MS/MS runs, determination of precursor ion m/z values and elution profiles, and the integration of mass chromatograms based on determined parameters for identified peptides. A total of 38 glycopeptides from 22 different proteins were significantly differentially abundant across the case/control pools (P<0.01, Student’s t test) and their abundances led to a near complete separation of case and control pools based on hierarchical clustering. The differential abundances of three of these candidate proteins were verified by commercially available ELISAs applied in the pools. Strong positive correlations between glycopeptide mass chromatograms and ELISA-measured protein abundance was observed for all of the selected glycoproteins.
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影响因子：
2.9
作者：
Hongsachart, Piyorot;Huang-Liu, Rosa;Chen, Shui-Tein
通讯作者：
Chen, Shui-Tein
影响因子：
3.9
作者：
Lindner, H;Helliger, W
通讯作者：
Helliger, W
影响因子：
7
作者：
Kuzyk, Michael A.;Smith, Derek;Borchers, Christoph H.
通讯作者：
Borchers, Christoph H.
影响因子：
3.3
作者：
Abbatiello, Susan E.;Pan, Yuan-Xiang;Conrads, Thomas P.
通讯作者：
Conrads, Thomas P.
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y