BACKGROUND: Anterior gradient 2 (AGR2) is a secreted, cancer-associated protein in many types of epithelia cancer cells; however current immunoassays lack sufficient sensitivity for its detection in biofluids. To address this issue, we developed an ultra-sensitive targeted mass spectrometric assay for quantification of AGR2 in urine and serum. METHODS: Selected reaction monitoring (SRM)-based AGR2 assay was configured based on two selected proteotypic peptides. Digested peptides from clinical samples were processed by a PRISM (high pressure and high resolution separations coupled with intelligent selection and multiplexing) strategy, which incorporates high pH reversed-phase LC separations to fractionate and select target fractions for subsequent LC-SRM analyses. RESULTS: The SRM assay for AGR2 showed a reproducibility of
Revised: March 24, 2014 |
Published: February 28, 2014
Citation
Shi T., Y. Gao, S. Quek, T.L. Fillmore, C.D. Nicora, D. Su, and R. Zhao, et al. 2014.A highly sensitive targeted mass spectrometric assay for quantification of low-abundance AGR2 in human urine and serum.Journal of Proteome Research 13, no. 2:875-882. PNWD-SA-10119. doi:10.1021/pr400912c