TY - JOUR
T1 - Analysis of serum total and free PSA using immunoaffinity depletion coupled to SRM
T2 - Correlation with clinical immunoassay tests
AU - Liu, Tao
AU - Hossain, Mahmud
AU - Schepmoes, Athena A.
AU - Fillmore, Thomas L.
AU - Sokoll, Lori J.
AU - Kronewitter, Scott R.
AU - Izmirlian, Grant
AU - Shi, Tujin
AU - Qian, Wei Jun
AU - Leach, Robin J.
AU - Thompson, Ian M.
AU - Chan, Daniel W.
AU - Smith, Richard D.
AU - Kagan, Jacob
AU - Srivastava, Sudhir
AU - Rodland, Karin D.
AU - Camp, David G.
N1 - Funding Information:
Portions of this work were supported by the National Cancer Institute Early Detection Research Network Interagency Agreement Y01-CN-05013-29 (to K.D.R. and D.G.C.) and National Institutes of Health Grants P41 RR018522 (to R.D.S.), U01 CA115102 (to D.W.C.), and U01 CA86402 and P30 CA054174-18 (to I.M.T). The experimental work described herein was performed in the Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by the Department of Energy and located at Pacific Northwest National Laboratory, which is operated by Battelle Memorial Institute for the Department of Energy under Contract DE-AC05-76RL0 1830 .
PY - 2012/8/3
Y1 - 2012/8/3
N2 - Recently, selected reaction monitoring mass spectrometry (SRM-MS) has been more frequently applied to measure low abundance biomarker candidates in tissues and biofluids, owing to its high sensitivity and specificity, simplicity of assay configuration, and exceptional 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. 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 good correlation in several independent clinical serum sample sets. These results demonstrate that the workflows and SRM assays developed here provide an attractive alternative for reliably measuring candidate biomarkers in human blood, without the need to develop affinity reagents. Furthermore, the simultaneous measurement of multiple biomarkers, including the free and bound forms of PSA, can be performed in a single multiplexed analysis using high-resolution liquid chromatographic separation coupled with SRM-MS. This article is part of a Special Issue entitled: Translational Proteomics.
AB - Recently, selected reaction monitoring mass spectrometry (SRM-MS) has been more frequently applied to measure low abundance biomarker candidates in tissues and biofluids, owing to its high sensitivity and specificity, simplicity of assay configuration, and exceptional 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. 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 good correlation in several independent clinical serum sample sets. These results demonstrate that the workflows and SRM assays developed here provide an attractive alternative for reliably measuring candidate biomarkers in human blood, without the need to develop affinity reagents. Furthermore, the simultaneous measurement of multiple biomarkers, including the free and bound forms of PSA, can be performed in a single multiplexed analysis using high-resolution liquid chromatographic separation coupled with SRM-MS. This article is part of a Special Issue entitled: Translational Proteomics.
KW - Free PSA
KW - Immunoaffinity depletion
KW - Immunoassay
KW - Selected reaction monitoring
KW - Serum
KW - Total PSA
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U2 - 10.1016/j.jprot.2012.01.035
DO - 10.1016/j.jprot.2012.01.035
M3 - Article
C2 - 22846433
AN - SCOPUS:84864082548
SN - 1874-3919
VL - 75
SP - 4747
EP - 4757
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 15
ER -