Impact of the depletion of high-abundance proteins in blood plasma/serum on the proteome of these media in growing farm animals

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.

Application of two-dimensional gel electrophoresis technique for protein profiling of Indian black gram varieties and detection of adulteration in black gram-based food products using comparative proteomics

“…1a, b and c). This might be due to the presence of high abundance acidic proteins (Lepczynski et al, 2014, Magdeldin et al, 2014). Low abundance proteins were found to be well separated with distinct identifiable spots.…”

“…4, samples (a)–(e) represent black gram papad (5), samples (f)–(h) represent instant medu vada mixes (3), (i)–(g) are the 2D patterns obtained for black gram flour (2) and (k) represents papad atta (1) . The region above MW 61.0 kDa showed horizontal streaking due to the presence of high-abundance acidic proteins (Lepczynski et al, 2014, Magdeldin et al, 2014). The 2D-GE pattern of these eleven samples were compared to patterns obtained for black gram varieties as well as refined flour and white pea (Fig.…”