Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods
Abstract
Background: The established methods for detecting prostate cancer (CaP) are based on tests using PSA (blood), PCA3 (urine), and AMACR (tissue) as biomarkers in patient samples. The demonstration of ERG oncoprotein overexpression due to gene fusion in CaP has thus provided ERG as an additional biomarker. Based on this, we hypothesized that ERG protein quantification methods can be of use in the diagnosis of prostate cancer. Methods: Therefore, an antibody-free assay for ERG3 protein detection was developed based on PRISM (high-pressure high-resolution separations with intelligent selection and multiplexing)-SRM (selected reaction monitoring) mass spectrometry. We utilized TMPRSS2-ERG positive VCaP and TMPRSS2-ERG negative LNCaP cells to simulate three different sample types (cells, tissue, and post-DRE urine sediment). Results: Recombinant ERG3 protein spiked into LNCaP cell lysates could be detected at levels as low as 20 pg by PRISM-SRM analysis. The sensitivity of the PRISM-SRM assay was around approximately 10,000 VCaP cells in a mixed cell population model of VCaP and LNCaP cells. Interestingly, ERG protein could be detected in as few as 600 VCaP cells spiked into female urine. The sensitivity of the in-house enzyme-linked immunosorbent assay (ELISA) was similar to the PRISM-SRM assay, with detection of 30 pg of purified recombinantmore »
- Authors:
- more »
- Publication Date:
- Research Org.:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1184918
- Report Number(s):
- PNNL-SA-107827
Journal ID: ISSN 1479-5876; 48505; 400412000
- Grant/Contract Number:
- AC05-76RL01830
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Translational Medicine
- Additional Journal Information:
- Journal Volume: 13; Journal Issue: 1; Journal ID: ISSN 1479-5876
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Environmental Molecular Sciences Laboratory
Citation Formats
He, Jintang, Schepmoes, Athena A., Shi, Tujin, Wu, Chaochao, Fillmore, Thomas L., Gao, Yuqian, Smith, Richard D., Qian, Weijun, Rodland, Karin D., Liu, Tao, Camp, David G., Rastogi, Anshu, Tan, Shyh-Han, Yan, Wusheng, Mohamed, Ahmed A., Huang, Wei, Banerjee, Sreedatta, Kagan, Jacob, Srivastava, Sudhir, McLeod, David, Srivastava, Shiv, Petrovics, Gyorgy, Dobi, Albert, and Srinivasan, Alagarsamy. Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods. United States: N. p., 2015.
Web. doi:10.1186/s12967-015-0418-z.
He, Jintang, Schepmoes, Athena A., Shi, Tujin, Wu, Chaochao, Fillmore, Thomas L., Gao, Yuqian, Smith, Richard D., Qian, Weijun, Rodland, Karin D., Liu, Tao, Camp, David G., Rastogi, Anshu, Tan, Shyh-Han, Yan, Wusheng, Mohamed, Ahmed A., Huang, Wei, Banerjee, Sreedatta, Kagan, Jacob, Srivastava, Sudhir, McLeod, David, Srivastava, Shiv, Petrovics, Gyorgy, Dobi, Albert, & Srinivasan, Alagarsamy. Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods. United States. https://doi.org/10.1186/s12967-015-0418-z
He, Jintang, Schepmoes, Athena A., Shi, Tujin, Wu, Chaochao, Fillmore, Thomas L., Gao, Yuqian, Smith, Richard D., Qian, Weijun, Rodland, Karin D., Liu, Tao, Camp, David G., Rastogi, Anshu, Tan, Shyh-Han, Yan, Wusheng, Mohamed, Ahmed A., Huang, Wei, Banerjee, Sreedatta, Kagan, Jacob, Srivastava, Sudhir, McLeod, David, Srivastava, Shiv, Petrovics, Gyorgy, Dobi, Albert, and Srinivasan, Alagarsamy. Thu .
"Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods". United States. https://doi.org/10.1186/s12967-015-0418-z. https://www.osti.gov/servlets/purl/1184918.
@article{osti_1184918,
title = {Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods},
author = {He, Jintang and Schepmoes, Athena A. and Shi, Tujin and Wu, Chaochao and Fillmore, Thomas L. and Gao, Yuqian and Smith, Richard D. and Qian, Weijun and Rodland, Karin D. and Liu, Tao and Camp, David G. and Rastogi, Anshu and Tan, Shyh-Han and Yan, Wusheng and Mohamed, Ahmed A. and Huang, Wei and Banerjee, Sreedatta and Kagan, Jacob and Srivastava, Sudhir and McLeod, David and Srivastava, Shiv and Petrovics, Gyorgy and Dobi, Albert and Srinivasan, Alagarsamy},
abstractNote = {Background: The established methods for detecting prostate cancer (CaP) are based on tests using PSA (blood), PCA3 (urine), and AMACR (tissue) as biomarkers in patient samples. The demonstration of ERG oncoprotein overexpression due to gene fusion in CaP has thus provided ERG as an additional biomarker. Based on this, we hypothesized that ERG protein quantification methods can be of use in the diagnosis of prostate cancer. Methods: Therefore, an antibody-free assay for ERG3 protein detection was developed based on PRISM (high-pressure high-resolution separations with intelligent selection and multiplexing)-SRM (selected reaction monitoring) mass spectrometry. We utilized TMPRSS2-ERG positive VCaP and TMPRSS2-ERG negative LNCaP cells to simulate three different sample types (cells, tissue, and post-DRE urine sediment). Results: Recombinant ERG3 protein spiked into LNCaP cell lysates could be detected at levels as low as 20 pg by PRISM-SRM analysis. The sensitivity of the PRISM-SRM assay was around approximately 10,000 VCaP cells in a mixed cell population model of VCaP and LNCaP cells. Interestingly, ERG protein could be detected in as few as 600 VCaP cells spiked into female urine. The sensitivity of the in-house enzyme-linked immunosorbent assay (ELISA) was similar to the PRISM-SRM assay, with detection of 30 pg of purified recombinant ERG3 protein and 10,000 VCaP cells. On the other hand, qRT-PCR exhibited a higher sensitivity, as TMPRSS2-ERG transcripts were detected in as few as 100 VCaP cells, in comparison to NanoString methodologies which detected ERG from 10,000 cells. Conclusions: Based on this data, we propose that the detection of both ERG transcriptional products with RNA-based assays, as well as protein products of ERG using PRISM-SRM assays, may be of clinical value in developing diagnostics and prognostics assays for prostate cancer given their sensitivity, specificity, and reproducibility.},
doi = {10.1186/s12967-015-0418-z},
journal = {Journal of Translational Medicine},
number = 1,
volume = 13,
place = {United States},
year = {Thu Jan 01 00:00:00 EST 2015},
month = {Thu Jan 01 00:00:00 EST 2015}
}
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Targeted Quantification of Low ng/mL Level Proteins in Human Serum without Immunoaffinity Depletion
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Direct multiplexed measurement of gene expression with color-coded probe pairs
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Common Gene Rearrangements in Prostate Cancer
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5′ UTR Control of Native ERG and of Tmprss2:ERG Variants Activity in Prostate Cancer
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Antibody-Based Detection of ERG Rearrangement-Positive Prostate Cancer
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The Present and Future of Prostate Cancer Urine Biomarkers
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Predominance of ERG-negative high-grade prostate cancers in African American men
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- Farrell, James; Young, Denise; Chen, Yongmei
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Antibody-free PRISM–SRM for multiplexed protein quantification: is this the new competition for immunoassays in bioanalysis?
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- Shi, Tujin; Qian, Wei-Jun
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Low Frequency of the ERG Oncogene Alterations in Prostate Cancer Patients from India
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