Antibody-based enrichment of peptides on magnetic beads for mass-spectrometry-based quantification of serum biomarkers.

ABSTRACT: A major bottleneck for validation of new clinical diagnostics is the development of highly sensitive and specific assays for quantifying proteins. We previously described a method, stable isotope standards with capture by antipeptide antibodies, wherein a specific tryptic peptide is selected as a stoichiometric representative of the protein from which it is cleaved, is enriched from biological samples using immobilized antibodies, and is quantitated using mass spectrometry against a spiked internal standard to yield a measure of protein concentration. In this study, we optimized a magnetic-bead-based platform amenable to high-throughput peptide capture and demonstrated that antibody capture followed by mass spectrometry can achieve ion signal enhancements on the order of 10(3), with precis

SUBMITTER: Whiteaker JR 

PROVIDER: S-EPMC1852426 | biostudies-literature | 2007 Mar

REPOSITORIES: biostudies-literature