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Microchip Electrophoresis with Mass Spectrometry Detection and Glycoprotein Analysis: Application in the Glycoproteomic Analysis of Trypanosoma Cruzi using Lectin Affinity

Grant number: 08/05648-0
Support type:Scholarships in Brazil - Post-Doctorate
Effective date (Start): September 01, 2008
Effective date (End): July 31, 2009
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
Principal researcher:Emanuel Carrilho
Grantee:Evandro Piccin
Home Institution: Instituto de Química de São Carlos (IQSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil

Abstract

This project involves the development of instrumentation in microchip electrophoresis with mass spectrometry detection (ME-MS) for glycoprotein analysis. ME-MS systems will be evaluated with two different electrospray (ESI) interfaces (sheath-flow and sheathless) by optimizing the analytical conditions for the separation and identification of glycopeptides derived from two model glycoproteins: RNase B and IgA.The ME-MS system (sheath-flow or sheathless) with the best analytical performance will be selected for a glycoproteomic study of the Trypanosoma cruzi parasite, which is the causative agent of Chagas' disease. The T. cruzi glycoproteins will be separated from the cellular lysate by using Lectin Affinity Chromatography (LAC) with two different lectins: concavalinA and jacalin. These lectins have specific selectivity to different polysaccharide structures and, therefore, they will select different kinds of glycoproteins.The ME-MS glycopeptide analyses will be compared with conventional CE-MS analysis, as CE-MS is a well established technique for glycoproteomic studies. The MS results obtained for the glycopeptides separated by ME and CE, as well as their fragmentation MS analysis (MS/MS), will be combined with bioinformatics for the identification of glycoproteins in the cellular membrane of T. cruzi.

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