Purpose To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases. Design Laboratory investigation. Methods Normal vitreous was obtained from eight human donor eyes and from eight eyes of patients undergoing diabetic vitrectomy. Vitreous specimens were subjected to SDS-PAGE and MALDI-TOF-MS analysis. Six specific antibodies were used to identify proteins using Western blot. Four proteins were localized within ocular tissue in a normal donor eyebank eye. Results We found eight distinct bands on SDS-PAGE in normal vitreous and two additional bands (hemoglobin) in eyes with diabetic retinopathy. Proteins were identified using MALDI-TOF-MS and confirmed by Western blot. We established a quantitative analysis of relative protein concentrations of undiluted vitreous. Immunohistochemistry localized selected proteins within the posterior segment layers. Conclusions We present a normal human vitreous protein profile using current technologies and provide a baseline for comparison to ocular disease states. Tissue distribution of vitreous proteins may help to elucidate more specific protein function.
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