Quantitative analysis of prostate specific antigen isoforms using immunoprecipitation and stable isotope labeling mass spectrometry

Yi Ting Chen, Li Ping Tuan, Hsiao Wei Chen, I. An Wei, Min Yuan Chou, Han Min Chen, Yu Chang Tyan, Sung-Fang Chen

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Prostate specific antigen (PSA) is a widely used serum marker for prostate cancer (PCa), but has limited specificity for distinguishing early PCa from benign prostatic hyperplasia (BPH). Recently, proPSAs, comprised of native proPSA, as well as truncated proPSA forms, [-2] proPSA, [-5] proPSA, and [-7] proPSA, have been shown to be better diagnostic targets than PSA for PCa. Stable isotope labeling-multiple reaction monitoring mass spectrometry (SIL/MRM-MS) has been frequently used to measure low-abundance biomarkers in tissues and biofluids, owing to its high sensitivity and specificity, simplicity, and multiplexing capability. In this study, we have developed and optimized a strategy using immunoprecipitation in conjunction with SIL/MRM-MS assay which is capable of sensitive and accurate quantification of proPSA in serum. Since serum and plasma are by far the most complex biological fluids, the immunoprecipitation workflow was optimized to achieve sufficient sensitivity, efficiencies of protein purification with immunoaffinity depletion were determined. The developed strategy can detect proPSA and PSA with a limit of detection (LOD) and limit of quantitation (LOQ) at nanogram per milliliter levels, corresponding to a concentration 6 orders-of-magnitude lower than the most abundant serum proteins. Furthermore, the simultaneous measurement of multiple biomarkers, including the mature and precursor forms of PSA, can be achieved in a single multiplexed analysis using LC/MRM-MS. The strategy demonstrated here provides an attractive alternative to ELISAs or RIAs for the reliably measurement of proPSA to improve the specificity of PCa diagnosis.

Original languageEnglish
Pages (from-to)545-553
Number of pages9
JournalAnalytical Chemistry
Volume87
Issue number1
DOIs
Publication statusPublished - 2015 Jan 6

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Isotope Labeling
Prostate-Specific Antigen
Immunoprecipitation
Isotopes
Labeling
Mass spectrometry
Mass Spectrometry
Prostatic Neoplasms
Protein Isoforms
Biomarkers
Chemical analysis
Workflow
Monitoring
Prostatic Hyperplasia
Serum
Multiplexing
Purification
Limit of Detection
Blood Proteins
Assays

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Quantitative analysis of prostate specific antigen isoforms using immunoprecipitation and stable isotope labeling mass spectrometry. / Chen, Yi Ting; Tuan, Li Ping; Chen, Hsiao Wei; Wei, I. An; Chou, Min Yuan; Chen, Han Min; Tyan, Yu Chang; Chen, Sung-Fang.

In: Analytical Chemistry, Vol. 87, No. 1, 06.01.2015, p. 545-553.

Research output: Contribution to journalArticle

Chen, Yi Ting ; Tuan, Li Ping ; Chen, Hsiao Wei ; Wei, I. An ; Chou, Min Yuan ; Chen, Han Min ; Tyan, Yu Chang ; Chen, Sung-Fang. / Quantitative analysis of prostate specific antigen isoforms using immunoprecipitation and stable isotope labeling mass spectrometry. In: Analytical Chemistry. 2015 ; Vol. 87, No. 1. pp. 545-553.
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