Quantitation of gla proteins by mass spectrometric analysis
AssigneesUniversity of Washington, Washington State University
InventorsBhagwat Prasad, Kenneth E. Thummel, Abdul Basit Shaikh, Allan E. Rettie
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Abstract
Methods of LC-MS/MS quantification of γ-carboxylated proteins in plasma, serum, or blood, including dried blood spots, are disclosed. The methods can be used to determine patient-specific dosing of anticoagulant drugs and diagnosis of liver diseases, such as hepatocellular carcinoma.
Claims20
A method for quantifying one or more of γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins in a biological sample, comprising:
- (a) extracting a sample comprising one or more of γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins from the biological sample having a first concentration, to provide a solution having a second concentration comprising one or more of γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins;
- (b) contacting the γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins in the solution having the second concentration with a protease, to provide a solution of peptides comprising one or more of γ-carboxylated peptides, γ-carboxylated peptide proteoforms, and des-carboxylated peptides;
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- (c) determining the quantity of the one or more of γ-carboxylated peptides, γ-carboxylated peptide proteoforms, and des-carboxylated peptides in the solution of peptides.
A method for quantifying one or more of γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins in a biological sample, comprising:
- (a) extracting a sample comprising one or more of γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins from the biological sample having a first concentration, to provide a solution having a second concentration comprising one or more of γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins;
- (b) contacting the γ-carboxylated proteins, γ-carboxylated protein proteoforms, and des-carboxylated proteins in the solution having the second concentration with a protease, to provide a solution of peptides comprising one or more of γ-carboxylated peptides, γ-carboxylated peptide proteoforms, and des-carboxylated peptides;
- (c) derivatizing the one or more of γ-carboxylated peptides, γ-carboxylated peptide proteoforms, and des-carboxylated peptides of the solution of peptides, to provide one or more derivatized γ-carboxylated peptides, derivatized γ-carboxylated peptide proteoforms, and derivatized des-carboxylated peptides;
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- (d) determining the quantity of the one or more of derivatized γ-carboxylated peptides, derivatized γ-carboxylated peptide proteoforms, and derivatized des-carboxylated peptides in the solution of peptides.
Dependencies are inferred from the claim text.