Method for protein expression analysis

Details Method for protein expression analysis Method for protein expression analysis

G - Physics

01

N

G01N 33/68 (2006.01)

Patent

CA 2471346

The present invention is a method for analysis of at least peptides comprising extracting proteins from at least one set of cells; digesting the extracted proteins; derivatising the protein fragment mixture with an isotopically labelled reagent molecule; separating the protein fragment mixture by multi- dimensional chromatography; and analysing the protein fragment mixture by mass spectroscopy (MS) in parent ion or neutral loss scanning mode, thereby detecting or measuring the amounts of the labelled protein fragments. In a preferred embodiment, two sets of cells are combined in the method in order to compare the expression levels of two different states. Moreover, the invention relates to a kit for use in the present labelling method.

L'invention concerne un procédé d'analyse d'au moins un peptide comprenant les étapes suivantes : extraire des protéines d'au moins un ensemble de cellules, traiter lesdites protéines extraites ; dérivatiser le mélange de fragments de protéines au moyen d'une molécule réactive marquée de manière isotope ; séparer ledit mélange de fragments de protéines par chromatographie multidimensionnelle ; et analyser le mélange de fragments de protéines par spectroscopie de masse (MS) en ion parent ou en mode par balayage de perte neutre, tout en détectant ou mesurant les quantités des fragments de protéines marquées. Dans un mode de réalisation préféré, deux ensembles de cellules sont combinés dans ledit procédé afin de comparer les niveaux d'expression de deux états différents. De plus, l'invention concerne un kit d'utilisation dans ledit procédé de marquage.

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