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Title: Facile carrier-assisted targeted mass spectrometric approach for proteomic analysis of low numbers of mammalian cells

Abstract

There is an unmet technical challenge for mass spectrometry (MS)-based proteomic analysis of single mammalian cells. Quantitative proteomic analysis of single cells has been previously achieved by antibody-based immunoassays but is limited by the availability of high-quality antibodies. Herein we report a facile targeted MS-based proteomics method, termed cPRISM-SRM (carrier-assisted high-pressure, high-resolution separations with intelligent selection and multiplexing coupled to selected reaction monitoring), for reliable analysis of low numbers of mammalian cells. The method capitalizes on using “carrier protein” to assist processing of low numbers of cells with minimal loss, high-resolution PRISM separation for target peptide enrichment, and sensitive SRM for protein quantification. We have demonstrated that cPRISM-SRM has sufficient sensitivity to quantify proteins expressed at ≥200,000 copies per cell at the single-cell level and ≥3000 copies per cell in 100 mammalian cells. We envision that with further improvement cPRISM-SRM has the potential to move toward targeted MS-based single-cell proteomics.

Authors:
 [1];  [1];  [1]; ORCiD logo [1];  [1];  [2]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]
  1. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  2. Central South Univ., Hunan (China)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1557087
Report Number(s):
[PNNL-SA-130600]
[Journal ID: ISSN 2399-3642]
Grant/Contract Number:  
[AC05-76RL01830]
Resource Type:
Accepted Manuscript
Journal Name:
Communications Biology
Additional Journal Information:
[ Journal Volume: 1; Journal Issue: 1]; Journal ID: ISSN 2399-3642
Publisher:
Springer Nature
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Shi, Tujin, Gaffrey, Matthew J., Fillmore, Thomas L., Nicora, Carrie D., Yi, Lian, Zhang, Pengfei, Shukla, Anil K., Wiley, H. Steven, Rodland, Karin D., Liu, Tao, Smith, Richard D., and Qian, Wei-Jun. Facile carrier-assisted targeted mass spectrometric approach for proteomic analysis of low numbers of mammalian cells. United States: N. p., 2018. Web. doi:10.1038/s42003-018-0107-6.
Shi, Tujin, Gaffrey, Matthew J., Fillmore, Thomas L., Nicora, Carrie D., Yi, Lian, Zhang, Pengfei, Shukla, Anil K., Wiley, H. Steven, Rodland, Karin D., Liu, Tao, Smith, Richard D., & Qian, Wei-Jun. Facile carrier-assisted targeted mass spectrometric approach for proteomic analysis of low numbers of mammalian cells. United States. doi:10.1038/s42003-018-0107-6.
Shi, Tujin, Gaffrey, Matthew J., Fillmore, Thomas L., Nicora, Carrie D., Yi, Lian, Zhang, Pengfei, Shukla, Anil K., Wiley, H. Steven, Rodland, Karin D., Liu, Tao, Smith, Richard D., and Qian, Wei-Jun. Mon . "Facile carrier-assisted targeted mass spectrometric approach for proteomic analysis of low numbers of mammalian cells". United States. doi:10.1038/s42003-018-0107-6. https://www.osti.gov/servlets/purl/1557087.
@article{osti_1557087,
title = {Facile carrier-assisted targeted mass spectrometric approach for proteomic analysis of low numbers of mammalian cells},
author = {Shi, Tujin and Gaffrey, Matthew J. and Fillmore, Thomas L. and Nicora, Carrie D. and Yi, Lian and Zhang, Pengfei and Shukla, Anil K. and Wiley, H. Steven and Rodland, Karin D. and Liu, Tao and Smith, Richard D. and Qian, Wei-Jun},
abstractNote = {There is an unmet technical challenge for mass spectrometry (MS)-based proteomic analysis of single mammalian cells. Quantitative proteomic analysis of single cells has been previously achieved by antibody-based immunoassays but is limited by the availability of high-quality antibodies. Herein we report a facile targeted MS-based proteomics method, termed cPRISM-SRM (carrier-assisted high-pressure, high-resolution separations with intelligent selection and multiplexing coupled to selected reaction monitoring), for reliable analysis of low numbers of mammalian cells. The method capitalizes on using “carrier protein” to assist processing of low numbers of cells with minimal loss, high-resolution PRISM separation for target peptide enrichment, and sensitive SRM for protein quantification. We have demonstrated that cPRISM-SRM has sufficient sensitivity to quantify proteins expressed at ≥200,000 copies per cell at the single-cell level and ≥3000 copies per cell in 100 mammalian cells. We envision that with further improvement cPRISM-SRM has the potential to move toward targeted MS-based single-cell proteomics.},
doi = {10.1038/s42003-018-0107-6},
journal = {Communications Biology},
number = [1],
volume = [1],
place = {United States},
year = {2018},
month = {8}
}

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