Proteomic sample preparation for blast wound characterization

Abstract

BackgroundBlast 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.ResultsTo 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.ConclusionHigh 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.

Wound fluid sampling methods for proteomic studies: A scoping review

“…Vacuum, drainage, or other external devices were used in 124 of the 280 (44%) studies and thus were the most frequently used methods of sample collection 22–27,29–146 ; the vast majority of sample collections using this method (75%, 103/138), were from surgical wounds (Figure 2B). Three other frequently used methods were extraction from absorbent material (31%, 86/280),…”

iTRAQ based investigation of plasma proteins in HIV infected and HIV/HBV coinfected patients — C9 and KLK are related to HIV/HBV coinfection

“…Subsequently, 14-high-abundant proteins from human biological fluids were greatly decreased by MARS Hu-14, which improved the resolution and dynamic range for proteomic analyses. This technology has been applied successfully for investigations of low abundance members of plasma/serum proteomes and has led to the discovery of biomarkers (Silbiger et al, 2011;Chromy et al, 2014;Johansen et al, 2009;Darde et al, 2007). The present data showed that some lower abundance protein bands were enriched after removing higher abundance proteins.…”

Wound outcome in combat injuries is associated with a unique set of protein biomarkers

“…All effluent samples were subjected to removal of high abundant proteins to improve overall spot clarity and separated by 2-D DIGE according to protocols established by Chromy et al [ 16 ]. Spot maps were analyzed by DeCyder 7.0 by which a total of 1800 spots were detected and quantified in each the 22 gels using the DIA module, and gel-to-gel spot matching was performed using the BVA module.…”