Diabetic retinopathy is a microvascular complication caused by diabetes and the leading cause of blindness in adults. Diabetic retinopathy has initiative stages such as non-proliferative diabetic retinopathy (NPDR), vision-threatening stages such as proliferative diabetic retinopathy (PDR), neovascularization occurring stages such as neovascuar glaucoma (NVG). Chronic hyperglycemia is a major cause of all microvascular complications derived from diabetes. However, it is difficult to predict diabetic retinopathy and its progression as the diagnostic biomarkers are not yet fully understood. In this study, we used quantitative proteomics to analyze AH (Aqueous Humor) proteins from patients with NPDR, PDR, and NVG. The aim of this study was to obtain a panel of AH proteins that were differentially expressed between NPDR and PDR and between PDR and NVG for the investigation of biomarkers and pathogenesis of the 3 serious complications. Totally about 2000 proteins were identified using a high-resolution orbitrap mass spectrometer coupled online to a nanoflow-LC system. Using both label-free quantification and TMT-based quantification, 44 differentially expressed proteins were enriched. We also validated total 49 proteins from profiling experiments and references using a triple-quadrupole mass spectrometer for quantification of targeted proteins.