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Se Joon Woo, Hye-Jung Kim, Eui Jin Suh, Ji Hyun Park, Jeeyun Ahn, Duck Jin Hwang, Ji Eun Lee, Kyu Hyung Park, Cheolju Lee; In-depth analysis of blood plasma proteome for discovery of age-related macular degeneration biomarkers. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4996.
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To find plasma biomarkers for age-related macular degeneration, we performed comprehensive proteomic analyses on blood plasma of AMD and age-matched healthy controls
We used a 4D protein profiling method on pooled plasma of patient and control groups. (1) Abundant protein depletion coupled with separation strategies - Gel-Eluted Liquid Fraction Entrapment Electrophoresis (GELFrEE) for protein and (2) Isoelectrofocusing of tryptic peptides by OFFGEL (3) Liquid chromatography and tandem mass analsys (4) LC-MS/MS. Global functional analyses and canonical pathway analyses were carried out using Ingenuity Pathway Analysis for proteins which were differentially expressed (more than 2- fold). Validation of candidate biomarkers was performed using ELISA and western blot on individual samples.
Using this profiling strategy, we identify 5042 unique peptides from 353 proteins, ranging over six orders of magnitude in abundance. Canonical pathways associated with AMD were identified for example, LXR/RXR activation, acute phase response signaling, complement cascades and actin cytoskeleton signaling. To develop potential biomarker proteins for AMD, we selected 7 proteins for further verification.
Our results implicate altered systemic pathways of AMD and provide a new protein database for hypothesis-driven and discovery based studies of AMD.
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