Proteome analysis of vitreous humor in retinal detachment using two different flow-charts for protein fractionation

- Two fractionation strategies were proposed for the vitreous proteome analysis in retinal detachment.
- A total of 127 proteins were identified in vitreous of patients with retinal detachment.
- From the proteins identified in vitreous, 68 had not yet been found in previous studies.
- Bidimensional fractionation is more suitable to increase the coverage of vitreous proteome.

The deeper understanding of retinal detachment (RD) pathogenesis may improve the visual outcome after surgery. Given the main role of the vitreous in retinal eye diseases, two strategies were explored to identify its proteome in RD. Fractionation techniques such as anion exchange chromatography (IEX) and SDS-PAGE combined with MALDI-TOF/TOF analysis allowed to identify 127 proteins in vitreous of RD patients. From these proteins, 19 were identified using only the IEX fractionation strategy, and 117 using a bidimensional (IEX and SDS-PAGE) fractionation. Of these proteins, 68 had not yet been found in other vitreous proteomic studies. The fractionation with IEX and SDS-PAGE largely improved the number of identified proteins proving that it is crucial to combine several methodologies to cover vitreous proteome.