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

Title: Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests

Abstract

Sandwich immunoassay is the standard technique used in clinical labs for quantifying protein biomarkers for disease detection, monitoring and therapeutic intervention. Albeit highly sensitive, the development of a specific immunoassay is rather time-consuming and associated with extremely high cost due to the requirement for paired immunoaffinity reagents of high specificity. Recently, mass spectrometry-based methods, specifically selected reaction monitoring mass spectrometry (SRM-MS), have been increasingly applied to measure low abundance biomarker candidates in tissue and biofluids, owing to high sensitivity and specificity, simplicity of assay configuration, and great multiplexing capability. In this study, we report for the first time the development of immunoaffinity depletion-based workflows and SRM-MS assays that enable sensitive and accurate quantification of total and free prostate-specific antigen (PSA) in serum without the requirement for specific PSA antibodies. With stable isotope dilution and external calibration, low ng/mL level detection of both total and free PSA was consistently achieved in both PSA-spiked female serum samples and actual patient serum samples. Moreover, comparison of the results obtained when SRM PSA assays and conventional immunoassays were applied to the same samples showed very good correlation (R2 values ranging from 0.90 to 0.99) in several independent clinical serum sample sets, including a setmore » of 33 samples assayed in a blinded test. These results demonstrate that the workflows and SRM assays developed here provide an attractive alternative for reliably measuring total and free PSA in human blood. Furthermore, simultaneous measurement of free and total PSA and many other biomarkers can be performed in a single analysis using high-resolution liquid chromatographic separation coupled with SRM-MS.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
1050795
Report Number(s):
PNNL-SA-83777
43796; 400412000; TRN: US201218%%1564
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Journal of Proteomics
Additional Journal Information:
Journal Volume: 75; Journal Issue: 15
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; 59 BASIC BIOLOGICAL SCIENCES; CALIBRATION STANDARDS; CORRELATIONS; DETECTION; DISEASES; IMMUNOASSAY; MASS SPECTROSCOPY; PROTEINS; STABLE ISOTOPES; Environmental Molecular Sciences Laboratory

Citation Formats

Liu, Tao, Hossain, Mahmud, Schepmoes, Athena A, Fillmore, Thomas L, Sokoll, Lori J, Kronewitter, Scott R, Izmirlian, Grant, Shi, Tujin, Qian, Weijun, Leach, Robin, Thompson, Ian M, Chan, Daniel W, Smith, Richard D, Kagan, Jacob, Srinivastava, Sudhir, Rodland, Karin D, and Camp, David G. Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests. United States: N. p., 2012. Web. doi:10.1016/j.jprot.2012.01.035.
Liu, Tao, Hossain, Mahmud, Schepmoes, Athena A, Fillmore, Thomas L, Sokoll, Lori J, Kronewitter, Scott R, Izmirlian, Grant, Shi, Tujin, Qian, Weijun, Leach, Robin, Thompson, Ian M, Chan, Daniel W, Smith, Richard D, Kagan, Jacob, Srinivastava, Sudhir, Rodland, Karin D, & Camp, David G. Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests. United States. https://doi.org/10.1016/j.jprot.2012.01.035
Liu, Tao, Hossain, Mahmud, Schepmoes, Athena A, Fillmore, Thomas L, Sokoll, Lori J, Kronewitter, Scott R, Izmirlian, Grant, Shi, Tujin, Qian, Weijun, Leach, Robin, Thompson, Ian M, Chan, Daniel W, Smith, Richard D, Kagan, Jacob, Srinivastava, Sudhir, Rodland, Karin D, and Camp, David G. 2012. "Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests". United States. https://doi.org/10.1016/j.jprot.2012.01.035.
@article{osti_1050795,
title = {Analysis of Serum Total and Free PSA Using Immunoaffinity Depletion Coupled to SRM: Correlation with Clinical Immunoassay Tests},
author = {Liu, Tao and Hossain, Mahmud and Schepmoes, Athena A and Fillmore, Thomas L and Sokoll, Lori J and Kronewitter, Scott R and Izmirlian, Grant and Shi, Tujin and Qian, Weijun and Leach, Robin and Thompson, Ian M and Chan, Daniel W and Smith, Richard D and Kagan, Jacob and Srinivastava, Sudhir and Rodland, Karin D and Camp, David G},
abstractNote = {Sandwich immunoassay is the standard technique used in clinical labs for quantifying protein biomarkers for disease detection, monitoring and therapeutic intervention. Albeit highly sensitive, the development of a specific immunoassay is rather time-consuming and associated with extremely high cost due to the requirement for paired immunoaffinity reagents of high specificity. Recently, mass spectrometry-based methods, specifically selected reaction monitoring mass spectrometry (SRM-MS), have been increasingly applied to measure low abundance biomarker candidates in tissue and biofluids, owing to high sensitivity and specificity, simplicity of assay configuration, and great multiplexing capability. In this study, we report for the first time the development of immunoaffinity depletion-based workflows and SRM-MS assays that enable sensitive and accurate quantification of total and free prostate-specific antigen (PSA) in serum without the requirement for specific PSA antibodies. With stable isotope dilution and external calibration, low ng/mL level detection of both total and free PSA was consistently achieved in both PSA-spiked female serum samples and actual patient serum samples. Moreover, comparison of the results obtained when SRM PSA assays and conventional immunoassays were applied to the same samples showed very good correlation (R2 values ranging from 0.90 to 0.99) in several independent clinical serum sample sets, including a set of 33 samples assayed in a blinded test. These results demonstrate that the workflows and SRM assays developed here provide an attractive alternative for reliably measuring total and free PSA in human blood. Furthermore, simultaneous measurement of free and total PSA and many other biomarkers can be performed in a single analysis using high-resolution liquid chromatographic separation coupled with SRM-MS.},
doi = {10.1016/j.jprot.2012.01.035},
url = {https://www.osti.gov/biblio/1050795}, journal = {Journal of Proteomics},
number = 15,
volume = 75,
place = {United States},
year = {Fri Aug 03 00:00:00 EDT 2012},
month = {Fri Aug 03 00:00:00 EDT 2012}
}