A highly sensitive targeted mass spectrometric assay for quantification of AGR2 protein in human urine and serum

Tujin Shi, Yuqian Gao, Sue Ing Quek, Thomas L. Fillmore, Carrie D. Nicora, Dian Su, Rui Zhao, Jacob Kagan, Sudhir Srivastava, Karin D. Rodland, Tao Liu, Richard D. Smith, Daniel W. Chan, David G. Camp, Alvin Y. Liu, Wei Jun Qian

Research output: Contribution to journalArticle

Abstract

Anterior gradient 2 (AGR2) is a secreted, cancer-associated protein in many types of epithelial cancer cells. We developed a highly sensitive targeted mass spectrometric assay for quantification of AGR2 in urine and serum. Digested peptides from clinical samples were processed by PRISM (high pressure and high resolution separations coupled with intelligent selection and multiplexing), which incorporates high pH reversed-phase liquid chromatography (LC) separations to fractionate and select target fractions for follow-on LC-selected reaction monitoring (LC-SRM) analyses. The PRISM-SRM assay for AGR2 showed a reproducibility of <10% CV and limit of quantification (LOQ) values of ∼130 pg/mL in serum and ∼10 pg per 100 μg of total protein mass in urine, respectively. A good correlation (R2 = 0.91) was observed for the measurable AGR2 concentrations in urine between SRM and enzyme-linked immunosorbent assay (ELISA). On the basis of an initial cohort of 37 subjects, urinary AGR2/PSA concentration ratios showed a significant difference (P = 0.026) between noncancer and cancer. Large clinical cohort studies are needed for the validation of AGR2 as a useful diagnostic biomarker for prostate cancer. Our work validated the approach of identifying candidate secreted protein biomarkers through genomics and measurement by targeted proteomics, especially for proteins where no immunoassays are available.

Original languageEnglish (US)
Pages (from-to)875-882
Number of pages8
JournalJournal of proteome research
Volume13
Issue number2
DOIs
StatePublished - Feb 7 2014

Keywords

  • AGR2
  • PRISM-SRM
  • PSA
  • human serum
  • human urine
  • prostate cancer

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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  • Cite this

    Shi, T., Gao, Y., Quek, S. I., Fillmore, T. L., Nicora, C. D., Su, D., Zhao, R., Kagan, J., Srivastava, S., Rodland, K. D., Liu, T., Smith, R. D., Chan, D. W., Camp, D. G., Liu, A. Y., & Qian, W. J. (2014). A highly sensitive targeted mass spectrometric assay for quantification of AGR2 protein in human urine and serum. Journal of proteome research, 13(2), 875-882. https://doi.org/10.1021/pr400912c