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

doi:10.1021/pr400912c

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

School of Medicine

Research output: Contribution to journal › Article › peer-review

45 Scopus citations

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 language	English (US)
Pages (from-to)	875-882
Number of pages	8
Journal	Journal of proteome research
Volume	13
Issue number	2
DOIs	https://doi.org/10.1021/pr400912c
State	Published - Feb 7 2014

Keywords

AGR2
PRISM-SRM
PSA
human serum
human urine
prostate cancer

ASJC Scopus subject areas

General Chemistry
Biochemistry

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10.1021/pr400912c

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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

Shi, T, Gao, Y, Quek, SI, Fillmore, TL, Nicora, CD, Su, D, Zhao, R, Kagan, J, Srivastava, S, Rodland, KD, Liu, T, Smith, RD, Chan, DW, Camp, DG, Liu, AY & Qian, WJ 2014, 'A highly sensitive targeted mass spectrometric assay for quantification of AGR2 protein in human urine and serum', Journal of proteome research, vol. 13, no. 2, pp. 875-882. https://doi.org/10.1021/pr400912c

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

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.",

keywords = "AGR2, PRISM-SRM, PSA, human serum, human urine, prostate cancer",

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

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AU - Shi, Tujin

AU - Gao, Yuqian

AU - Quek, Sue Ing

AU - Fillmore, Thomas L.

AU - Nicora, Carrie D.

AU - Su, Dian

AU - Zhao, Rui

AU - Kagan, Jacob

AU - Srivastava, Sudhir

AU - Rodland, Karin D.

AU - Liu, Tao

AU - Smith, Richard D.

AU - Chan, Daniel W.

AU - Camp, David G.

AU - Liu, Alvin Y.

AU - Qian, Wei Jun

PY - 2014/2/7

Y1 - 2014/2/7

N2 - 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.

AB - 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.

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KW - PSA

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KW - human urine

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

Abstract

Keywords

ASJC Scopus subject areas

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