skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Mass Spectrometry-based Assay for High Throughput and High Sensitivity Biomarker Verification

Abstract

Searching for disease specific biomarkers has become a major undertaking in the biomedical research field as the effective diagnosis, prognosis and treatment of many complex human diseases are largely determined by the availability and the quality of the biomarkers. A successful biomarker as an indicator to a specific biological or pathological process is usually selected from a large group of candidates by a strict verification and validation process. To be clinically useful, the validated biomarkers must be detectable and quantifiable by the selected testing techniques in their related tissues or body fluids. Due to its easy accessibility, protein biomarkers would ideally be identified in blood plasma or serum. However, most disease related protein biomarkers in blood exist at very low concentrations (<1ng/mL) and are “masked” by many none significant species at orders of magnitude higher concentrations. The extreme requirements of measurement sensitivity, dynamic range and specificity make the method development extremely challenging. The current clinical protein biomarker measurement primarily relies on antibody based immunoassays, such as ELISA. Although the technique is sensitive and highly specific, the development of high quality protein antibody is both expensive and time consuming. The limited capability of assay multiplexing also makes the measurement an extremelymore » low throughput one rendering it impractical when hundreds to thousands potential biomarkers need to be quantitatively measured across multiple samples. Mass spectrometry (MS)-based assays have recently shown to be a viable alternative for high throughput and quantitative candidate protein biomarker verification. Among them, the triple quadrupole MS based assay is the most promising one. When it is coupled with liquid chromatography (LC) separation and electrospray ionization (ESI) source, a triple quadrupole mass spectrometer operating in a special selected reaction monitoring (SRM) mode, also known as multiple reaction monitoring (MRM), is capable of quantitatively measuring hundreds of candidate protein biomarkers from a relevant clinical sample in a single analysis. The specificity, reproducibility and sensitivity could be as good as ELISA. Furthermore, SRM MS can also quantify protein isoforms and post-translational modifications, for which traditional antibody-based immunoassays often don’t exist.« less

Authors:
;
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
1378050
Report Number(s):
PNNL-SA-105981
47301; 400412000
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Book
Resource Relation:
Related Information: Protein Analysis Using Mass Spectrometry: Accelerating Protein Biotherapeutics from Lab to Patient
Country of Publication:
United States
Language:
English
Subject:
Environmental Molecular Sciences Laboratory

Citation Formats

Guo, Xuejiang, and Tang, Keqi. Mass Spectrometry-based Assay for High Throughput and High Sensitivity Biomarker Verification. United States: N. p., 2017. Web. doi:10.1002/9781119371779.ch9.
Guo, Xuejiang, & Tang, Keqi. Mass Spectrometry-based Assay for High Throughput and High Sensitivity Biomarker Verification. United States. doi:10.1002/9781119371779.ch9.
Guo, Xuejiang, and Tang, Keqi. Wed . "Mass Spectrometry-based Assay for High Throughput and High Sensitivity Biomarker Verification". United States. doi:10.1002/9781119371779.ch9.
@article{osti_1378050,
title = {Mass Spectrometry-based Assay for High Throughput and High Sensitivity Biomarker Verification},
author = {Guo, Xuejiang and Tang, Keqi},
abstractNote = {Searching for disease specific biomarkers has become a major undertaking in the biomedical research field as the effective diagnosis, prognosis and treatment of many complex human diseases are largely determined by the availability and the quality of the biomarkers. A successful biomarker as an indicator to a specific biological or pathological process is usually selected from a large group of candidates by a strict verification and validation process. To be clinically useful, the validated biomarkers must be detectable and quantifiable by the selected testing techniques in their related tissues or body fluids. Due to its easy accessibility, protein biomarkers would ideally be identified in blood plasma or serum. However, most disease related protein biomarkers in blood exist at very low concentrations (<1ng/mL) and are “masked” by many none significant species at orders of magnitude higher concentrations. The extreme requirements of measurement sensitivity, dynamic range and specificity make the method development extremely challenging. The current clinical protein biomarker measurement primarily relies on antibody based immunoassays, such as ELISA. Although the technique is sensitive and highly specific, the development of high quality protein antibody is both expensive and time consuming. The limited capability of assay multiplexing also makes the measurement an extremely low throughput one rendering it impractical when hundreds to thousands potential biomarkers need to be quantitatively measured across multiple samples. Mass spectrometry (MS)-based assays have recently shown to be a viable alternative for high throughput and quantitative candidate protein biomarker verification. Among them, the triple quadrupole MS based assay is the most promising one. When it is coupled with liquid chromatography (LC) separation and electrospray ionization (ESI) source, a triple quadrupole mass spectrometer operating in a special selected reaction monitoring (SRM) mode, also known as multiple reaction monitoring (MRM), is capable of quantitatively measuring hundreds of candidate protein biomarkers from a relevant clinical sample in a single analysis. The specificity, reproducibility and sensitivity could be as good as ELISA. Furthermore, SRM MS can also quantify protein isoforms and post-translational modifications, for which traditional antibody-based immunoassays often don’t exist.},
doi = {10.1002/9781119371779.ch9},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2017},
month = {6}
}

Book:
Other availability
Please see Document Availability for additional information on obtaining the full-text document. Library patrons may search WorldCat to identify libraries that hold this book.

Save / Share:

Works referenced in this record:

Enhanced Sensitivity for Selected Reaction Monitoring Mass Spectrometry-based Targeted Proteomics Using a Dual Stage Electrodynamic Ion Funnel Interface
journal, April 2010

  • Hossain, Mahmud; Kaleta, David T.; Robinson, Errol W.
  • Molecular & Cellular Proteomics, Vol. 10, Issue 2
  • DOI: 10.1074/mcp.M000062-MCP201

Subambient Pressure Ionization with Nanoelectrospray Source and Interface for Improved Sensitivity in Mass Spectrometry
journal, March 2008

  • Page, Jason S.; Tang, Keqi; Kelly, Ryan T.
  • Analytical Chemistry, Vol. 80, Issue 5
  • DOI: 10.1021/ac702354b

Long-Gradient Separations Coupled with Selected Reaction Monitoring for Highly Sensitive, Large Scale Targeted Protein Quantification in a Single Analysis
journal, September 2013

  • Shi, Tujin; Fillmore, Thomas L.; Gao, Yuqian
  • Analytical Chemistry, Vol. 85, Issue 19
  • DOI: 10.1021/ac402105s

Electrospray ionization for mass spectrometry of large biomolecules
journal, October 1989


Chemically Etched Open Tubular and Monolithic Emitters for Nanoelectrospray Ionization Mass Spectrometry
journal, November 2006

  • Kelly, Ryan T.; Page, Jason S.; Luo, Quanzhou
  • Analytical Chemistry, Vol. 78, Issue 22, p. 7796-7801
  • DOI: 10.1021/ac061133r

Mass Spectrometric Quantitation of Peptides and Proteins Using Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA)
journal, April 2004

  • Anderson, N. Leigh; Anderson, Norman G.; Haines, Lee R.
  • Journal of Proteome Research, Vol. 3, Issue 2
  • DOI: 10.1021/pr034086h

Molecular Beams of Macroions
journal, September 1968

  • Dole, Malcolm; Mack, L. L.; Hines, R. L.
  • The Journal of Chemical Physics, Vol. 49, Issue 5
  • DOI: 10.1063/1.1670391

On the evaporation of small ions from charged droplets
journal, January 1976

  • Iribarne, J. V.
  • The Journal of Chemical Physics, Vol. 64, Issue 6
  • DOI: 10.1063/1.432536

A Highly Sensitive Targeted Mass Spectrometric Assay for Quantification of AGR2 Protein in Human Urine and Serum
journal, December 2013

  • Shi, Tujin; Gao, Yuqian; Quek, Sue Ing
  • Journal of Proteome Research, Vol. 13, Issue 2
  • DOI: 10.1021/pr400912c

Achieving 50% Ionization Efficiency in Subambient Pressure Ionization with Nanoelectrospray
journal, November 2010

  • Marginean, Ioan; Page, Jason S.; Tolmachev, Aleksey V.
  • Analytical Chemistry, Vol. 82, Issue 22
  • DOI: 10.1021/ac1019123

Analytical Properties of the Nanoelectrospray Ion Source
journal, January 1996

  • Wilm, Matthias; Mann, Matthias
  • Analytical Chemistry, Vol. 68, Issue 1
  • DOI: 10.1021/ac9509519

Ultrasensitive Sample Quantitation via Selected Reaction Monitoring Using CITP/CZE–ESI-Triple Quadrupole MS
journal, November 2012

  • Wang, Chenchen; Lee, Cheng S.; Smith, Richard D.
  • Analytical Chemistry, Vol. 84, Issue 23
  • DOI: 10.1021/ac302616m

Targeted Quantification of Low ng/mL Level Proteins in Human Serum without Immunoaffinity Depletion
journal, June 2013

  • Shi, Tujin; Sun, Xuefei; Gao, Yuqian
  • Journal of Proteome Research, Vol. 12, Issue 7
  • DOI: 10.1021/pr400178v

Antibody-free, targeted mass-spectrometric approach for quantification of proteins at low picogram per milliliter levels in human plasma/serum
journal, September 2012

  • Shi, T.; Fillmore, T. L.; Sun, X.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 38
  • DOI: 10.1073/pnas.1204366109