Comparative proteome analysis of peripheral blood mononuclear cells in systemic lupus erythematosus with iTRAQ quantitative proteomics

Abstract

To identify and quantify protein profiles from peripheral blood mononuclear cells (PBMC) of systemic lupus erythematosus (SLE) patients with isobaric Tagging for Relative and Absolute protein Quantification (iTRAQ)-based proteomic technology and to find differentially expressed proteins in SLE. PBMC were collected from patients of six stable SLE, six active SLE, six rheumatoid arthritis (RA), and six healthy donors. After protein extraction and concentration, the pooled protein content was labeled with iTRAQ reagents and then subjected to multiple chromatographic fractionation and tandem mass spectrometry. ProteinPilot™ 3.0 software and a database of IPI (International Protein Index) human 3.62 were used for database searching and statistical analysis. A total of 452 proteins were identified. Of these, 67 unique proteins were observed twofold or more alteration in levels across groups. The proteins determined support existing knowledge and uncover novel biomarker candidates. These results indicate that iTRAQ-based technology can serve as a useful aid for identification and quantification proteins from PBMC.

Identification of potential microRNA–target pairs associated with osteopetrosis by deep sequencing, iTRAQ proteomics and bioinformatics

“…Therefore, mass tagging of peptides ensures any differences in the protein expression levels are because of initial differences in the samples, and not an artifact in the sample handling or processing. 22 As a result, we revealed a large-scale proteomic changes in osteopetrosis. One protein, such as RhoA (IPI01015923), has been implicated to be involved in osteopetrosis.…”

Proteomic analysis of human plasma and peripheral blood mononuclear cells in Systemic Lupus Erythematosus patients

“…From the present literature [33][34][35][36][37][38][39], we note the importance of proteomics, which can help to find new therapeutic targets and it also could help to better understand the cellular mechanisms. …”

Comparative proteomic analysis of renal tissue in IgA nephropathy with iTRAQ quantitative proteomics

“…The MS/MS data from all 10 fractions were combined and subsequently analyzed using the Paragon Algorithm search engine and Human v3.62 downloaded from the EBI website (http://www. ebi.ac) (13).…”

“…The peptides in these fractions were then separated on a Tempo™LC Nanoflow and matrix-assisted laser desorption/ionization (MALDI) spotting system equipped with a reversed-phase Magic C18AQ coluIgAN. Each chromatography run yielded ~380 MALDI spots on a stainless steel MALDI target plate Agilent 1290 Infinity (2D-LC) (Santa Clara, CA, USA) (13). MS data were obtained using an Applied Biosystems 4800 Plus MALDI TOF/TOF.…”