June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
In-depth analysis of blood plasma proteome for discovery of age-related macular degeneration biomarkers
Author Affiliations & Notes
  • Se Joon Woo
    Ophthalmology, Seoul National University College of Medicine, Seoul, Republic of Korea
    Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea
  • Hye-Jung Kim
    Theragnosis Research Center, Korea Institute of Science and Technology, Seoul, Republic of Korea
  • Eui Jin Suh
    Theragnosis Research Center, Korea Institute of Science and Technology, Seoul, Republic of Korea
  • Ji Hyun Park
    Ophthalmology, Seoul National University College of Medicine, Seoul, Republic of Korea
    Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea
  • Jeeyun Ahn
    Ophthalmology, Seoul National University College of Medicine, Seoul, Republic of Korea
    Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea
  • Duck Jin Hwang
    Ophthalmology, Seoul National University College of Medicine, Seoul, Republic of Korea
    Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea
  • Ji Eun Lee
    Theragnosis Research Center, Korea Institute of Science and Technology, Seoul, Republic of Korea
  • Kyu Hyung Park
    Ophthalmology, Seoul National University College of Medicine, Seoul, Republic of Korea
    Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea
  • Cheolju Lee
    Theragnosis Research Center, Korea Institute of Science and Technology, Seoul, Republic of Korea
    Biomolecular Science, University of Science and Technology, Daejeon, Republic of Korea
  • Footnotes
    Commercial Relationships Se Joon Woo, None; Hye-Jung Kim, None; Eui Jin Suh, None; Ji Hyun Park, None; Jeeyun Ahn, None; Duck Jin Hwang, None; Ji Eun Lee, None; Kyu Hyung Park, None; Cheolju Lee, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 4996. doi:
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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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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: To find plasma biomarkers for age-related macular degeneration, we performed comprehensive proteomic analyses on blood plasma of AMD and age-matched healthy controls

Methods: 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.

Results: 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.

Conclusions: Our results implicate altered systemic pathways of AMD and provide a new protein database for hypothesis-driven and discovery based studies of AMD.

Keywords: 412 age-related macular degeneration • 663 proteomics  
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