"Product ion monitoring" assay for prostate-specific antigen in serum using a linear ion-trap

Journal of Proteome Research
Vathany KulasingamEleftherios P Diamandis

Abstract

While numerous strategies exist for biomarker discovery, the bottleneck to product development and routine use at the clinic is in the verification phase of candidate biomarkers. The aim of this study was to establish a robust and high-throughput product ion monitoring (PIM) assay that is potentially capable of rapidly verifying candidates from discovery phase experiments. Using prostate-specific antigen (PSA), a model biomarker, and a routinely used mass spectrometer for discovery platforms, an ion trap (LTQ, Thermo), the utility of this instrument to perform PIM was explored. The proteotypic doubly charged intact peptide LSEPAELTDAVK ( m/ z 637) fragmenting to m/ z 943 (PAELTDAVK) was monitored. A limit of detection of 10 attomoles with a coefficient of variation (CV) of <20% was obtained for a purified recombinant PSA digest. Immunoextraction of endogenous PSA from serum using a monoclonal antibody on a 96-well microtiter plate, followed by PIM on the LTQ, enabled quantification of PSA down to less than 1 ng/mL with a limit of detection of 0.1 ng/mL and CVs < 20%. Mascot searching and ion ratio confirmation further supported the conclusion that the quantified moiety in serum was the PSA peptide. We conclude that this methodo...Continue Reading

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Citations

Aug 8, 2012·Analytical and Bioanalytical Chemistry·Aurora CuoghiAldo Tomasi
Dec 18, 2008·Molecular & Cellular Proteomics : MCP·Uros KuzmanovEleftherios P Diamandis
May 15, 2010·Molecular & Cellular Proteomics : MCP·Johannes A HewelAndrew Emili
Mar 13, 2009·Briefings in Functional Genomics & Proteomics·Anastasia K Yocum, Arul M Chinnaiyan
Feb 9, 2013·BMC Medicine·Uros KuzmanovEleftherios P Diamandis
Mar 28, 2009·Clinical Chemistry·Jesse C SeegmillerRajiv Kumar
May 24, 2012·Expert Review of Molecular Diagnostics·Jérôme LemoineGeneviève Choquet-Kastylevsky
Jan 1, 2010·Expert Opinion on Medical Diagnostics·Joanna M Hunter, Eustache Paramithiotis
Sep 16, 2014·BMC Medicine·Eleftherios P Diamandis
Mar 19, 2013·Clinical Biochemistry·Maria D PasicEleftherios P Diamandis
Apr 3, 2010·Trends in Biotechnology·Pierre LescuyerDenis F Hochstrasser
Mar 6, 2016·Analytical Biochemistry·Angelique Florentinus-Mefailoski, John G Marshall
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Sep 3, 2020·Cells·Anna IllianoAngela Amoresano

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