Automated High-throughput Analysis of Proteins/Peptides in Blood-based Matrices Combining Immunoaffinity Purification and Ultra-fast SPE/MS/MS

摘要

Automated, high-throughput immunoaffinity extraction coupled with ultra - fast SPE/MS/MS allows for the rapid analysis of FLAG-peptideC in rat serum. Total analysis time from serum to results is 1 hr for 96 samples. The immunoaffinity cartridges were optimized for efficient use of capture antibody and could be reused multiple times. Immunoaffinity purification showed equivalent results between samples prepared in PBS or serum suggesting that recovery efficiency was unaffected by the sample matrix. The quantitation of FLAG-peptideC across a range of 7.8 to 250 nM produced good linearity, accuracy and precision results. This methodology may be useful for the analysis of similar biomolecules in complex blood-based matrices.