Dergiler / Turkish Journal of Veterinary and Animal Sciences / 2014 / Cilt: 38 - Sayı: 5
Impact of the depletion of high-abundance proteins in blood plasma/serum on the proteome of these media in growing farm animals
- Sayfa
- 490–495
- DOI
- —
Özet
The aim of the study was to use hexapeptide combinatorial libraries to evaluate the influence of high-abundance protein depletion on low-abundance protein enrichment, protein concentration, and 2D gel quality in plasma and serum proteomes of growing piglets and calves. Depleted and nondepleted samples of plasma/serum were resolved using 2D electrophoresis (2DE). Bioinformatic analysis of the 2D gels showed an increased number of detected spots on each gel after depletion of excess high-abundance proteins in calf plasma (+26.46%) and in piglet serum (+49.40%). Relative expression of selected low-abundance protein spots in the blood plasma/serum samples was higher after high-abundance protein depletion. Both media showed decreased expressions of high-abundance protein spot clusters on 2D gels after treatment. The results of this study confirm both the high efficiency of high-abundance protein depletion and the increase in concentrations of low-abundance proteins on 2D gels after treatment. The method of depletion is reproducible and it may be used as an alternative to immunodepletion for preparing blood plasma/serum samples from young, growing animals for 2DE. Increased content of the low-abundance proteins on the 2D gels enables complete analysis of plasma/serum proteome and thereby allows for the discovery of novel biomarkers that might be useful in veterinary medicine.
Abstract
The aim of the study was to use hexapeptide combinatorial libraries to evaluate the influence of high-abundance protein depletion on low-abundance protein enrichment, protein concentration, and 2D gel quality in plasma and serum proteomes of growing piglets and calves. Depleted and nondepleted samples of plasma/serum were resolved using 2D electrophoresis (2DE). Bioinformatic analysis of the 2D gels showed an increased number of detected spots on each gel after depletion of excess high-abundance proteins in calf plasma (+26.46%) and in piglet serum (+49.40%). Relative expression of selected low-abundance protein spots in the blood plasma/serum samples was higher after high-abundance protein depletion. Both media showed decreased expressions of high-abundance protein spot clusters on 2D gels after treatment. The results of this study confirm both the high efficiency of high-abundance protein depletion and the increase in concentrations of low-abundance proteins on 2D gels after treatment. The method of depletion is reproducible and it may be used as an alternative to immunodepletion for preparing blood plasma/serum samples from young, growing animals for 2DE. Increased content of the low-abundance proteins on the 2D gels enables complete analysis of plasma/serum proteome and thereby allows for the discovery of novel biomarkers that might be useful in veterinary medicine.