Proteomic Analyses of Vitreous in Proliferative Diabetic Retinopathy: Prior Studies and Future Outlook

Abstract

Vitreous fluid is becoming an increasingly popular medium for the study of retinal disease. Numerous studies have demonstrated that proteomic analysis of the vitreous from patients with proliferative diabetic retinopathy yields valuable molecular information regarding known and novel proteins and pathways involved in this disease. However, there is no standardized methodology for vitreous proteomic studies. Here, we share a suggested protocol for such studies and outline the various experimental and analytic methods that are currently available. We also review prior mass spectrometry-based proteomic studies of the vitreous from patients with proliferative diabetic retinopathy, discuss common pitfalls of these studies, and propose next steps for moving the field forward.

Keywords: bioinformatics; mass spectrometry; proliferative diabetic retinopathy; proteomics; vitreous.

Figure 1

Suggested workflow for sample validation and processing. Protein concentration and sample integrity should be assessed prior to analysis. Depending on the investigators’ goals, EVs and hemoglobin may be quantified prior to further processing. Abundant proteins should be depleted and concentration should be reassessed to ensure sufficient depth of coverage. SDS-PAGE can then be repeated to validate prior steps before proceeding with MS.