SELDI-TOF-MS ProteinChip Array profiling of tears from patients with dry eye
SELDI-TOF-MS ProteinChip Array profiling of tears from patients with dry eye
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DOI：
10.1167/iovs.04-0448
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发表时间：
2005-03-01
影响因子：
4.4
通讯作者：
Pfeiffer, N
中科院分区：
文献类型：
--
作者：
Grus, FH;Podust, VN;Pfeiffer, N
目的.泪液中的蛋白质和多肽在眼表疾病中起重要作用。在以前的研究中，干眼症患者的电泳蛋白质谱已经显示出变化。本工作的目的是确定表面增强激光解吸/电离飞行时间质谱（SELDI-TOF-MS）蛋白芯片阵列（Ciphergen Biosystems，Inc.，Fremont，CA）技术用于泪液中蛋白质和肽的自动分析。对干眼症患者（DRY，n = 88）和健康受试者（CTRL，n = 71）进行了检查。使用SELDI-TOF-MS ProteinChip Arrays分析他们的泪液蛋白，该蛋白具有通过实验室液体处理机器人工作站制备的三种不同的色谱表面（CM 10阳离子交换，Q10阴离子交换和H50反相）。通过多元统计技术和人工神经网络对数据进行分析，最重要的生物标志物通过串联MS纯化和鉴定。检测到泪液蛋白和肽的复杂模式。不同的色谱表面揭示了脂质运载蛋白和溶菌酶等蛋白质的选择性富集。判别分析显示干眼症患者的蛋白质谱有非常显著的变化（P < 0.001）。通过七肽多标记物面板，人工神经网络可以区分干眼症患者和健康个体，特异性和灵敏度为90%。生物标志物的鉴定表明，干眼症患者的炎症标志物增加，而一些可能具有保护功能的蛋白质减少。结论。SELDI-TOF-MS技术似乎非常适合于泪液中肽和蛋白质的大规模筛选。这种高灵敏度的方法大大缩短了分析时间，并提供了具有很高质量精度的蛋白质图谱。因此，它可能成为一个非常有用的工具，在寻找潜在的生物标志物的诊断和新的治疗方法，在眼部疾病，如干眼。
PURPOSE. Protein and peptides in tears play an important role in ocular surface diseases. In previous studies, changes have been demonstrated in the electrophoretic protein profiles of patients with dry eye. The purpose of this work was to determine the usefulness of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) ProteinChip Array (Ciphergen Biosystems, Inc., Fremont, CA) technology for the automated analysis of proteins and peptides in tear fluid.METHODS. Patients with dry eye (DRY, n = 88) and healthy subjects (CTRL, n = 71) were examined. Their tear proteins were analyzed using SELDI-TOF-MS ProteinChip Arrays with three different chromatographic surfaces (CM10 cation exchange, Q10 anion exchange, and H50 reversed-phase) prepared by means of a laboratory liquid-handling robotic work-station. The data were analyzed by multivariate statistical techniques and artificial neural networks, and the most important biomarkers were purified and identified by tandem MS.RESULTS. Complex patterns of tear proteins and peptides were detected. The different chromatographic surfaces revealed the selective enrichment of proteins such as lipocalin and lysozyme. Discriminant analysis demonstrated highly significant changes in the protein profiles in patients with dry eye (P < 0.001). With a seven-peptide multimarker panel, an artificial neural network could differentiate between patients with dry eye and healthy individuals with a specificity and sensitivity of 90%. The identification of biomarkers revealed an increase of inflammatory markers in patients with dry eye and a decrease of some proteins that may have protective functions.CONCLUSIONS. The SELDI-TOF-MS technology seems to be ideally suitable for the mass screening of peptides and proteins in tears. This highly sensitive approach dramatically reduces the analysis time and provides protein profiles with great mass accuracy. Thus, it may become a very useful tool in the search for potential biomarkers for diagnosis and new therapeutics in ocular diseases such as dry eye.