Journal
PROTEOMICS CLINICAL APPLICATIONS
Volume 2, Issue 9, Pages 1265-1280Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/prca.200800017
Keywords
cluster analysis; marker proteins; quantitative proteomics; two-dimensional gel; vitreoretinal diseases
Funding
- NIH [EY13877, EY12190, RR17703]
- NATIONAL CENTER FOR RESEARCH RESOURCES [P20RR017703] Funding Source: NIH RePORTER
- NATIONAL EYE INSTITUTE [P30EY012190, R01EY013877] Funding Source: NIH RePORTER
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Vitreous samples collected in retinopathic surgeries have diverse properties, making proteomics analysis difficult. We report a cluster analysis to evade this difficulty. Vitreous and subretinal fluid samples were collected from 60 patients during surgical operation of non-proliferative diabetic retinopathy, proliferative diabetic retinopathy, proliferative vitreoretinopathy, and rhegmatogenous retinal detachment. For controls, we collected vitreous fluid from patients of idiopathic macular hole, epiretinal, and from a healthy postmortem donor. Proteins from these samples were subjected to quantitative proteomics using two-dimensional gel electrophoresis. We selected 105 proteins robustly expressed among ca. 400 protein spots and subjected them to permutation test. By using permutation test analysis we observed unique variations in the expression of some of these proteins in vitreoretinal diseases when compared to the control and to each other: (i) the levels of inflammation -associated proteins such as alpha 1 -antitrypsin, apolipoprotein A4, albumin, and transferrin were significantly higher in all four types of vitreoretinal diseases, and (ii) each vitreoretinal disease elevated a unique set of proteins, which can be interpreted based on the pathology of retinopathy. Our protocol will be effective for the study of protein expression in other types of clinical samples of diverse properties.
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