[en] In shotgun proteomics, the gold standard technique is reversed-phase liquid chromatography coupledto mass spectrometry. Many researches have been carried out to study the effects on identification per-formances of chromatographic parameters such as the stationary phase and column dimensions, mobilephase composition and flow rate, as well as the gradient slope and length. However, little attention isusually paid to the injection solvent composition.In this study, we investigated the effect of the injection solvent on protein identification parameters(number of distinct peptides, amino acid coverage and MS/MS search score) as well as sensitivity. Trypticpeptides from six different proteins, covering a wide range of physicochemical properties, were employedas training set. Design of experiments was employed as a tool to highlight the factors related to thecomposition of the injection solvent that significantly influenced the obtained results. Optimal resultsfor the training set were applied to analysis of more complex samples. The experiments pointed outoptimising the composition of the injection solvent had a strong beneficial effect on all the consideredresponses. On the basis of these results, an approach to determine optimal conditions was proposed tomaximise the protein identification performances and detection sensitivity.
Research Center/Unit :
Centre Interfacultaire de Recherche du Médicament - CIRM
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Houbart, Virginie ; Université de Liège > Département de pharmacie > Analyse des médicaments
COBRAIVILLE, Gaël ; Centre Hospitalier Universitaire de Liège - CHU > Service de rhumatologie
Nys, Gwenaël ; Université de Liège > Département de pharmacie > Analyse des médicaments
Merville, Marie-Paule ; Université de Liège > Département de pharmacie > Chimie médicale
Fillet, Marianne ; Université de Liège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
Impact of injection solvent composition on protein identification incolumn-switching chip-liquid chromatography/mass spectrometry
Publication date :
2016
Journal title :
Journal of Chromatography. A
ISSN :
0021-9673
eISSN :
1873-3778
Publisher :
Elsevier Science, Amsterdam, Netherlands
Volume :
1445
Pages :
27-35
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
Service public de Wallonie : Direction générale opérationnelle de l'économie, de l'emploi et de la recherche - DG06 F.R.S.-FNRS - Fonds de la Recherche Scientifique
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