Amino acid analysis based on isotope Amino acid analysis based on isotope-dilution and high-resolution mass spectrometry for sensitive protein quantification.

摘要

The AAA-MS method displays several advantages that makes it especially suited for the quantification of isotope-dilution proteomicsstandards: 1. VERSATILITY: PSAQ proteins or AQUA peptides can be hydrolyzed and quantified easily. 2. SPEED: 3 to 5 hours are necessary for a quantification analysis, 10 samples can be processed in paralled. 3. SENSITIVITY: only 5 pmoles of PSAQ (250ng of a 45kDa protein) or 250 pmoles of AQUA standards (375ng of 1500Da peptide) are needed for quantification. Thus, AAA-MS is 400 fold more sensitive than classical AAA assays! 4. ACCURACY: A good correlation was obtained between AAA-MS and classical AAA results regarding PSAQ standards quantification and AQUA peptides.