Optimization of An Immunoaffinity LC-SRM MS Assay to Quantita te Recombinant Human Alpha-1 Antitrypsin (rhAAT) from Biological Matrices

摘要

1. A highly specific translatable LC-SRM MS assay has been developed to measure recombinant and endogenous human AAT protein from serum and BAL matrices using surrogate peptide analysis. 2. The immunoprecipitation step was optimized to a 2-hr sample preparation method followed by an overnight digest with trypsin. 3. An Agilent 6490 LC-MS/MS operating in SRM mode was used for sample quantitation with an LOD of 1 ng/mL from 50 uL of sample. 4. This fit for purpose method has been qualified for mouse serum and BAL fluid and is expected to translate to the clinic.