ORBi: De Bock Muriel - Comparison of three methods for fractionation and enrichment of low molecular weight proteins for SELDI-TOF-MS differential analysis

Reference : Comparison of three methods for fractionation and enrichment of low molecular weight pro...


	Document type :	Scientific journals : Article
	Discipline(s) :	Human health sciences : Pharmacy, pharmacology & toxicology
	To cite this reference:	http://hdl.handle.net/2268/8787

Title :	Comparison of three methods for fractionation and enrichment of low molecular weight proteins for SELDI-TOF-MS differential analysis
Language :	English
Author, co-author :	De Bock, Muriel [Université de Liège - ULg > > > GIGA - Membres]
	De Seny, Dominique [Université de Liège - ULg > > Rhumatologie >]
	Meuwis, Marie-Alice [Université de Liège - ULg > > GIGA-Management : Plate-forme protéomique >]
	Servais, Anne-Catherine [Université de Liège - ULg > Département de pharmacie > Analyse des médicaments >]
	Minh, Tran Quang [Affiland > > > >]
	Closset, Jean [Université de Liège - ULg > Département des sciences biomédicales et précliniques > Département des sciences biomédicales et précliniques >]
	Chapelle, Jean-Paul [Université de Liège - ULg > Département de pharmacie > Chimie médicale >]
	Louis, Edouard [Université de Liège - ULg > Département des sciences cliniques > Hépato-gastroentérologie - Relations académiques et scientifiques (Médecine) >]
	Malaise, Michel [Université de Liège - ULg > Département des sciences cliniques > Rhumatologie >]
	Merville, Marie-Paule [Université de Liège - ULg > Département de pharmacie > Chimie médicale >]
	Fillet, Marianne [Université de Liège - ULg > Département de pharmacie > Analyse des médicaments >]
Publication date :	2010
Journal title :	Talanta
Publisher :	Elsevier Science
Volume :	82
Pages :	245-254
Audience :	International
ISSN :	0039-9140
e-ISSN :	1873-3573
City :	Amsterdam
Country :	The Netherlands
Keywords :	[en] Protein prefractionation ; Major protein depletion ; Plasma ; Serum ; SELDI-TOF-MS
Abstract :	[en] In most diseases, the clinical need for serum/plasma markers has never been so crucial, not only for diagnosis, but also for the selection of the most efficient therapies, as well as exclusion of ineffective or toxic treatment. Due to the high sample complexity, prefractionation is essential for exploring the deep proteome and finding specific markers. In this study, three different sample preparation methods (i.e., highly abundant protein precipitation, restricted access materials (RAM) combined with IMAC chromatography and peptide ligand affinity beads) were investigated in order to select the best fractionation step for further differential proteomic experiments focusing on the LMW proteome (MW inferior to 40,000 Da). Indeed, the aim was not to cover the entire plasma/serum proteome, but to enrich potentially interesting tissue leakage proteins. These three methods were evaluated on their reproducibility, on the SELDI-TOF-MS peptide/protein peaks generated after fractionation and on the information supplied. The studied methods appeared to give complementary information and presented good reproducibility (below 20%). Peptide ligand affinity beads were found to provide efficient depletion of HMW proteins and peak enrichment in protein/peptide profiles.
Funders :	Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS ; Fonds Léon Fredericq ; Centre Anti-Cancéreux ; Télévie
Permalink :	http://hdl.handle.net/2268/8787

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