Figure 1.

Study design for comparative proteomics based mass spectrometry for identifying candidate biomarkers in AMD. 200 μg pooled samples were reduced and alkylated by using Dithiothreitol and Iodoacetamide, respectively. The samples were digested overnight with trypsin (Promega) (1:20) at 37°C. Peptides from each group were labelled using 8plex tandem mass tags with technical replicates as per manufacturer's protocol (Catalogue # 90110 Thermo Fischer Scientific). Peptides derived from control were labelled by 126 and 127N, Early AMD was labelled by 127C and 128N, CNVM were labelled by 128C and 129N, GA was labelled by 129C and 130N and are pooled. The fractions were analyzed using Orbitrap Fusion mass spectrometer and the Raw data was analysed using Proteome discoverer software. The data was further validated using ELISA