Proteomic sample preparation for blast wound characterization

Brett A. Chromy*, Angela Eldridge, Jonathan A. Forsberg, Trevor S. Brown, Benjamin C. Kirkup, Eric Elster, Paul Luciw

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Background: Blast wounds often involve diverse tissue types and require substantial time and treatment for appropriate healing. Some of these subsequent wounds become colonized with bacteria requiring a better understanding of how the host responds to these bacteria and what proteomic factors contribute wound healing outcome. In addition, using reliable and effective proteomic sample preparation procedures can lead to novel biomarkers for improved diagnosis and therapy.Results: To address this need, suitable sample preparation for 2-D DIGE proteomic characterization of wound effluent and serum samples from combat-wounded patients was investigated. Initial evaluation of crude effluent and serum proved the necessity of high abundant protein depletion. Subsequently, both samples were successfully depleted using Agilent Multiple Affinity Removal system and showed greatly improved 2-D spot maps, comprising 1,800 and 1,200 protein spots, respectively.Conclusion: High abundant protein removal was necessary for both wound effluent and serum. This is the first study to show a successful method for high abundant protein depletion from wound effluent which is compatible with downstream 2-D DIGE analysis. This development allows for improved biomarker discovery in wound effluent and serum samples.

Original languageEnglish
Article number10
JournalProteome Science
Volume12
Issue number1
DOIs
StatePublished - 14 Feb 2014
Externally publishedYes

Keywords

  • 2-D DIGE
  • Biomarker discovery
  • Blast wound
  • High abundant protein removal
  • Proteomics
  • Serum
  • Wound effluent

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