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JIOMICS | VOL 5 | ISSUE 2 | DECEMBER 2015 | 1-62 JOURNAL OF INTEGRATED OMICS Journal of Integrated Omics A METHODOLOGICAL JOURNAL HTTP://WWW.JIOMICS.COM Special Issue: Proceeding Abstracts of the 4 th International Congress on Analytical Proteomics (ICAP 2015) Protein expression profiles of serum samples bipolar disorder patients mapped by 2-D DIGE coupled with nanoLC-MS/MS J.R. Jesus* 1 , R. M. Galazzi 1 , B. K. Campos 1 , A. Sussulini 1 , C. E. M. Banzatto 2 , J. L. C. Martínez 3 , M. A. Z. Arruda 1 1 Spectrometry Sample Preparation and Mechanization Group – GEPAM. Institute of Chemistry. University of Campinas -UNICAMP. 13083- 970. Campinas, SP. Brazil. 2 Department of Psychiatry. Faculty of Medical Science. University of Campinas - UNICAMP. 13083-970. Campinas, SP.Brazil. 3 Bioscope group. Chemistry Department. Faculty of Sciences and Technology. New University of Lisbon. Campus de Caparica. 2829 -516. Caparica. Portugal. *Corresponding author: jemmyson.jesus@iqm.unicamp.br Available Online: 31 December 2015 Abstract Purpose: Bipolar Disorder (BD), which affects up to 3% of the worldwide population, is a mental disease with unclear connections to other similar disorder. This study had aim application approach comparative proteomic to identify potential biomarkers in patient serum samples with bipolar disorder. Experimental description: This study was approved by the Ethics Committee of the Hospital das Clinicas (University of Campinas, Brazil). Fifty six serum samples (23-57 years old) were collected and classified into five groups: (A) healthy family control (n= 3), (B) healthy nofamily control (n= 9), (C) BD patients under treatment with lithium (Li) (n= 14), (D) schizophrenia patients (n= 25) and (E) other diagnostic patients under treatment with Li (n=4). The participants did not have other concomitant diseases such as cancer, endocrinological or metabolic diseases. Abundant proteins (IgG and albumin) depletion were performed using ProteoMiner spin columns. The analysis was performed using two-dimensional fluorescence difference-in-gel electrophoresis (2-D DIGE) coupled with nanoliquid chromatography-tandem mass spectrometry (nanoLC-MS/MS). For the analysis of 2-D DIGE, the samples were compared. As follows: (i) group A vs group B; (ii) group A vs group E; (iii) group B vs group E; (iv) group C vs group E; (v) group E vs group D. Results: Approximately 50 μg [(pH range 4-70) determined from the 2-D Quant Kit] of each sample and the corresponding amount of the internal pooled standard were labeled with 400 ρmol of CyDye DIGE Fluors minimal dyes. The ratios of protein abundance that increased or decreased more than 2.0 fold (t-test, p
JIOMICS | VOL 5 | ISSUE 2 | DECEMBER 2015 | 1-62 JOURNAL OF INTEGRATED OMICS Journal of Integrated Omics A METHODOLOGICAL JOURNAL HTTP://WWW.JIOMICS.COM Special Issue: Proceeding Abstracts of the 4 th International Congress on Analytical Proteomics (ICAP 2015) Western blotting - detection of a specific protein in MCF7 and MCF7-Z tissue J. Ner-Kluza* 1 , A. Drabik 1 , Michael Kubbutat 2 , Andreas Lingnau 2 , P. Suder 1,5 , G. Schroeder 4 , J. Silberring 1,3 1 Department of Biochemistry and Neurobiology., Faculty of Materials Sciences and Ceramics. AGH University of Science and Technology. Mickiewicza 30 Ave. 3 0-059 Krakow. Poland. 2 ProQinase GmbH. Freiburg, Germany. 3 Center of Polymer and Carbon Materials. Polish Academy of Sciences. Marii Skłodowskiej-Curie 34., 41-819 Zabrze. Poland. 4 Department of Supramolecular Chemistry. Faculty of Chemistry. Adam Mickiewicz University. Umultowska 89b. 61-614 Poznan. Poland. 5 AGH University of Science and Technology. Academic Centre for Materials and Nanotechnology. Mickiewicza 30. 30–059, Kraków, Poland. *Corresponding author: nerjoanna@gmail.com Available Online: 31 December 2015 Abstract Purpose: In the contemporary world, breast cancer is one of the most common and dangerous types of cancer in females. Comparison between two different variants of MCF-7 (MCF-7 vs MCF-7-Z) human breast cancer cell lines used in subcutaneous xenograft tumor models, has shown different tumor growth characteristics in mice. Glycosylated proteins are among post-translational modifications one of the most important types of proteins involved in tumor progression. The aim of the research was to search for the presence of selected glycoproteins, based on quantitative and qualitative analysis in cancer progression. Experimental description: For samples preparation we used the strategy utilizing western blotting and nanoLC-MS/MS analysis. After isolation of proteins from the tissue, glycoproteins were isolated by lectin affinity chromatography. Glycoproteins fractions were separated by the sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE), followed by two complementary analyses. One part of the gel was transferred onto PVDF membrane, and another part was CBB stained. Based on the images obtained after immunostaining, the bands were excised from the CBB-stained gel and analyzed by tandem mass spectrometry coupled to liquid chromatography (LC-MS/MS). Results: Using LC-MS/MS technique we have succeeded to identify 3000 differentially expressed proteins between MCF7 and MCF7-Z. The selected proteins were associated with cancer metastasis, which plays a vital role in tumor progression, and are involved in cell structure organization. The presence of the selected proteins was confirmed using western blotting. Conclusions: We have identified the potential biomarkers, which can be important in cancer progression. Keywords: cell lines MCF-7, glycoproteins, tumor, breast cancer. Acknowledgements: The research was supported by the grant EuroNanoMed "META" 05/EuroNanoMed/2012. 1-62: 50
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Volume 5 | Issue 2 | December 2015
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Randen Patterson Center for Computa
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Biodiversity Research Center, Acade
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Belgium Bart De Spiegeleer Ghent Un
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Instituto Superior Técnico, Centro
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Veronica Mainini Dept. Health Scien
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Xuequn Chen Department of Molecular
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Committees Conference Chairs 4 th I
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