two-dimensional electrophoresis Archives - Critical Care Science (CCS)

BACKGROUND AND OBJECTIVES: The diagnostic and treatment of sepsis continue to challenger all, and, more specific forms to approach are absolutely necessary. The objective of this study was to use proteomics techniques, two-dimensional electrophoresis and mass spectrometry, to verify the differential protein expression between serum of patients with sepsis and health controls. METHODS: Samples of serum the 30 patients with sepsis, caused for different types of microorganisms and serum of 30 health controls were obtained for analysis. Next, were submitted to 2D-SDS-PAGE, gels compared, selection of spots for excision and digestion with trypsin, being the peptides analyzed for MALDI TOF-TOF. The obtained spectrums were processed (Mascot-matrix science) for protein identification in NCBInr Data Bank. RESULTS: Image analyses showed several spots with differential expressions in the gels of the patients with sepsis in relation to the controls. The protein identification of some of these spots founded: Orosomucoid 1 precursor, Apolipoprotein A-IV, Apolipoprotein A-IV precursor, Haptoglobin protein precursor, Haptoglobin, Zinc finger protein, Serum amyloid A-1, Transthyretin, Nebulin, Complement C4, Alpha1-Antitrypsin, Unnamed protein product and others. CONCLUSIONS: Serum of the patients with different types of sepsis express characteristic protein profiles by 2D-SDS-PAGE compared with controls. The most expressed were from acute phase proteins and lipoproteins. It is possible in the future, with proteomics, create diagnostic panel of proteins, finding news biomarkers and targets for therapeutic interventions in sepsis. This is a first description, with proteomics, of the alterations in protein expression, in serum of the patients with sepsis.