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A. Wirthlin, N. Pfeiffer, F.H. Grus; ANALYSIS OF TEAR FLUID PROTEINS IN DRY–EYE BY PROTEIN–CHIPS: A NEW NON–INVASIVE APPROACH FOR BIOMARKER DETECTION . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3456.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose. The diagnosis of dry eye syndrome is commonly based on clinical tests such as the basal secretory test (BST). However, this test has no good correlation with the course of the disease and with other clinical tests available. The quantitative analysis of tear proteins could be useful for the diagnosis and follow–up of dry eye and other ocular surface diseases. The aim of this study was to analyze the tear protein pattern by means of a new ProteinChip approach which is very suitable for mass screen applications. Methods. A total of 233 eyes were examined in this study (DRY: n=127; CTRL: n=106). The patients were primarily grouped according to the results of the basic secretory test (BST). The tear proteins were analyzed by high–throughput surface–enhanced laser desorption ionization (SELDI) mass spectrometry proteomic technology based on ProteinChip arrays. The micro–scale design of the arrays allows the analysis of very small quantities of proteins. Three different protein chip surfaces reflecting different chromatographic separations were used: cationic exchanger arrays (CM–10), hydrophobic arrays (H50), and Cu–affinity chromatography (IMAC) under several binding conditions. The arrays were read by mass spectrometry and subsequently analyzed by multivariate statistics (e.g., analysis of discriminance). Results. Complex protein patterns could be demonstrated using all surfaces. The main protein peaks (e.g. lysozyme, lipocalin etc.) could be identified. In average more than 60 proteins could be detected on each surface. The analysis of discriminance could found a significant difference (P<0.001) between the protein profiles of dry–eye and normal tears. Furthermore, the SELDI technology was able to find complex protein and peptide profiles downto 2 kDa, which were not accessible in previous studies using electrophoretical techniques. Conclusion. The results show significant differences between the two groups. Our study demonstrates that biomarker profiles of tear proteins can be consistently and reproducibly used to differentiate between dry eye and normal tears. The SELDI technology might be a very promising technique for very fast and reliable mass screening of tear protein samples, which combines the sensitivity of two–dimensional electrophoresis with the mass accuracy of mass spectrometry analysis. Therefore, the analysis of biomarker profiles in tears will play an important role in the diagnosis, follow–up and clinical studies to assess the efficacy of treatment in dry eye and other ocular surface diseases.
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