Proteome analysis of vitreous humor in retinal detachment using two different flow-charts for protein fractionation

J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Sep 1:1061-1062:334-341. doi: 10.1016/j.jchromb.2017.07.049. Epub 2017 Jul 29.

Abstract

The deeper understanding of retinal detachment (RD) pathogenesis may improve the visual outcome after surgery. Given the main role of the vitreous in retinal eye diseases, two strategies were explored to identify its proteome in RD. Fractionation techniques such as anion exchange chromatography (IEX) and SDS-PAGE combined with MALDI-TOF/TOF analysis allowed to identify 127 proteins in vitreous of RD patients. From these proteins, 19 were identified using only the IEX fractionation strategy, and 117 using a bidimensional (IEX and SDS-PAGE) fractionation. Of these proteins, 68 had not yet been found in other vitreous proteomic studies. The fractionation with IEX and SDS-PAGE largely improved the number of identified proteins proving that it is crucial to combine several methodologies to cover vitreous proteome.

Keywords: Electrophoresis; Liquid chromatography; Mass spectrometry; Proteomic analysis; Retinal detachment; Vitreous humor.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Eye Proteins / analysis
  • Eye Proteins / chemistry
  • Female
  • Humans
  • Male
  • Middle Aged
  • Peptide Fragments / analysis
  • Peptide Fragments / chemistry
  • Proteome / analysis*
  • Proteome / chemistry
  • Proteomics / methods*
  • Retinal Detachment / metabolism*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Vitreous Body / chemistry*

Substances

  • Eye Proteins
  • Peptide Fragments
  • Proteome