Isobaric tags for relative and absolute quantification-based quantitative proteomic analysis for identification of potential serum biomarkers for lower limb arteriosclerosis obliterans

Abstract

Objective Using a proteomic approach we aimed to determine potential serum biomarkers for diagnosis and managememt of lower limb arteriosclerosis obliterans (LASO).Methods Serum samples were collected from 23 patients with LASO and 23 healthy volunteer subjects.Isobaric tags for relative and absolute quantitation (iTRAQ)-tagging combined with two-dimensional liquid chromatographytandem masss pectrometry (2D LC-MS/MS) analysis was used to identification of differentially expressed proteins between LASO patients and healthy subjects.Candidates were validated in independent cohorts of LASO patients and healthy subjects by Westerm blotting.Results Using a proteomic approach,we identified 20 serum proteins are differentially expressed in LASO patients as compared with that in healthy subjects,of which 12 proteins increased more than two-fold,while 8 proteins decreased more than two-fold among the LASO patients.Western blotting test verified that Olfactomedin-1 (OLFM1) was up-regulated in LASO group as compared with the control group (2.05 ±0.66 vs 0.96 ±0.18,P ＜0.01).Conclusion Using an iTRAQ MS approach,we identification of differentially expressed proteins between LASO patients and healthy subjects,which might serve as potential candidates for diagnostic or therapeutic measures. Key words: Lower limb arteriosclerosis obliterans ; Proteomic ; Serum ; Biomarkers