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      Proteomic Analysis of Human Serum from Patients with Chronic Kidney Disease

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          Abstract

          Chronic kidney disease (CKD) is an important public health problem in the world. The aim of our research was to identify novel potential serum biomarkers of renal injury. ELISA assay showed that cytokines and chemokines IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, Eotaxin, FGFb, G-CSF, GM-CSF, IP-10, MCP-1, MIP-1α, MIP-1β, PDGF-1bb, RANTES, TNF-α and VEGF were significantly higher (R > 0.6, p value < 0.05) in the serum of patients with CKD compared to healthy subjects, and they were positively correlated with well-established markers (urea and creatinine). The multiple reaction monitoring (MRM) quantification method revealed that levels of HSP90B2, AAT, IGSF22, CUL5, PKCE, APOA4, APOE, APOA1, CCDC171, CCDC43, VIL1, Antigen KI-67, NKRF, APPBP2, CAPRI and most complement system proteins were increased in serum of CKD patients compared to the healthy group. Among complement system proteins, the C8G subunit was significantly decreased three-fold in patients with CKD. However, only AAT and HSP90B2 were positively correlated with well-established markers and, therefore, could be proposed as potential biomarkers for CKD.

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          Most cited references65

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          Difference gel electrophoresis: a single gel method for detecting changes in protein extracts.

          We describe a modification of two-dimensional (2-D) polyacrylamide gel electrophoresis that requires only a single gel to reproducibly detect differences between two protein samples. This was accomplished by fluorescently tagging the two samples with two different dyes, running them on the same 2-D gel, post-run fluorescence imaging of the gel into two images, and superimposing the images. The amine reactive dyes were designed to insure that proteins common to both samples have the same relative mobility regardless of the dye used to tag them. Thus, this technique, called difference gel electrophoresis (DIGE), circumvents the need to compare several 2-D gels. DIGE is reproducible, sensitive, and can detect an exogenous difference between two Drosophila embryo extracts at nanogram levels. Moreover, an inducible protein from E. coli was detected after 15 min of induction and identified using DIGE preparatively.
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            Silver staining of proteins in polyacrylamide gels.

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              Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity.

              Mass spectrometry is a powerful technique for the identification of proteins at nanogram quantities. However, some degree of sample preparation prior to mass spectrometry is required, and silver-stained protein gel samples are most problematic. Here we report our strategy to obtain peptide mass profiles from silver-stained protein gel samples from one- or two-dimensional gels by destaining prior to enzymatic digestion. This study demonstrates that by using the destaining method, the sensitivity and quality of mass spectra is increased for matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometric analysis, permitting more proteins to be identified by peptide mass database analysis.
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                Author and article information

                Journal
                Biomolecules
                Biomolecules
                biomolecules
                Biomolecules
                MDPI
                2218-273X
                07 February 2020
                February 2020
                : 10
                : 2
                : 257
                Affiliations
                [1 ]Institute of Fundamental Medicine and Biology, Kazan Federal University, 420008 Kazan, Tartastan, Russia; alexander.laikov@ 123456yandex.ru (A.L.); mimarkelova@ 123456gmail.com (M.M.); nazyrova.95@ 123456yandex.ru (R.K.); rizvanov@ 123456gmail.com (A.R.)
                [2 ]Republican Clinical Hospital Ministry of Health Republic of Tatarstan, 420064 Kazan, Tatarstan, Russia; alfiz.mar82@ 123456inbox.ru (A.M.); milyash@ 123456inbox.ru (M.H.)
                [3 ]Department of Urology and Nephrology, Kazan State Medical Academy, 420012 Kazan, Tatarstan, Russia
                [4 ]Department of Microbiology and Immunology, University of Nevada, Reno, NV 89557, USA; sv.khaiboullina@ 123456gmail.com
                Author notes
                [* ]Correspondence: magnolina@ 123456list.ru (Y.R.); sal.ilnur@ 123456gmail.com (I.S.); Tel.: +7-927-418-90-02 (Y.R.); +7-917-867-43-60 (I.S.)
                Author information
                https://orcid.org/0000-0003-1272-6100
                https://orcid.org/0000-0002-9427-5739
                Article
                biomolecules-10-00257
                10.3390/biom10020257
                7072325
                32046176
                96bedace-2d7b-41ef-b826-07cbe9fe63b0
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 21 January 2020
                : 05 February 2020
                Categories
                Article

                chronic kidney disease,inflammation,cytokines,biomarkers,2d-dige,multiple reaction monitoring

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