Protein fractionation and relative quantitation using PF 2D and ITRAQ for biomarker quest

0329307 - ÚŽFG 2010 RIV CZ eng K - Conference Paper (Czech conference)
Gadher, S. J. - Skalníková, Helena - Halada, Petr - Řehulka, Pavel - Chmelík, Josef - Kovářová, Hana
Protein fractionation and relative quantitation using PF 2D and ITRAQ for biomarker quest.
1st Central and Eastern European Proteomic Conference and 3rd Czech Proteomic Conference. Mělník: ÚŽFG, 2007, s. 7-8.
[1st Central and Eastern European proteomic Conference and 3rd Czech Proteomic Conference. Praha (CZ), 29.10.2007-31.10.2007]
R&D Projects: GA ČR GA301/05/0418; GA MŠMT LC07017
Institutional research plan: CEZ:AV0Z50450515; CEZ:AV0Z40310501; CEZ:AV0Z50200510
Keywords : protein fractionation * proteomics
Subject RIV: FD - Oncology ; Hematology

ProteomeLab PF 2D System - Protein Fractionation in 2 Dimensions (Beckman Coulter, Fullerton, CA, USA) has been developed to fractionate complex protein mixtures by chromatofocusing in the first dimension followed by high-resolution non-porous silica reversed phase chromatography (RP LC) in the second dimension. Despite the high-resolution power of ProteomeLab PF 2D, UV-based quantitation could be compromised due to possible co-elution of several proteins into one fraction. Hence, we present an optimized protocol for application of isobaric tags for relative and absolute quantitation (iTRAQ) and MALDI-TOF/TOF mass spectrometry to obtain quantitative data from peptides derived by tryptic digestions of intact proteins fractionated by ProteomeLab PF 2D technique. To demonstrate the feasibility of such an approach, protein expression patterns obtained from the ProteomeLab PF 2D fractionation of human T-lymphoblastic leukemia CEM cell line were utilised.
Permanent Link: http://hdl.handle.net/11104/0175379