Fifth-generation digital immunoassay for prostate-specific antigen by single molecule array technology

David H. Wilson, David W. Hanlon, Gail K. Provuncher, Lei Chang, Linan Song, Purvish P. Patel, Evan P. Ferrell, Herbert Lepor, Alan Wayne Partin, Daniel Wan-Yui Chan, Lori J Sokoll, Carol D. Cheli, Robert P. Thiel, David R. Fournier, David C. Duffy

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Measurement of prostate-specific antigen (PSA) in prostate cancer patients following radical prostatectomy (RP) has been hindered by the limit of quantification of available assays. Because radical prostatectomy removes the tissue responsible for PSA production, postsurgical PSA is typically undetectable with current assay methods. Evidence suggests, however, that more sensitive determination of PSA status following RP could improve assessment of patient prognosis and response to treatment and better target secondary therapy for those who may benefit most. We developed an investigational digital immunoassay with a limit of quantification 2 logs lower than current ultrasensitive third-generation PSA assays. METHODS: We developed reagents for a bead-based ELISA for use with high-density arrays of femtoliter-volume wells. Anti-PSA capture beads with immunocomplexes and associated enzyme labels were singulated within the wells of the arrays and interrogated for the presence of enzymatic product. We characterized analytical performance, compared its accuracy with a commercially available test, and analyzed longitudinal serum samples from a pilot study of 33 RP patients. RESULTS: The assay exhibited a functional sensitivity (20% interassay CV)

Original languageEnglish (US)
Pages (from-to)1712-1721
Number of pages10
JournalClinical Chemistry
Volume57
Issue number12
DOIs
StatePublished - Dec 2011

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Prostate-Specific Antigen
Immunoassay
Prostatectomy
Technology
Molecules
Assays
Labels
Prostatic Neoplasms
Enzyme-Linked Immunosorbent Assay
Tissue
Enzymes
Therapeutics
Serum

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Wilson, D. H., Hanlon, D. W., Provuncher, G. K., Chang, L., Song, L., Patel, P. P., ... Duffy, D. C. (2011). Fifth-generation digital immunoassay for prostate-specific antigen by single molecule array technology. Clinical Chemistry, 57(12), 1712-1721. https://doi.org/10.1373/clinchem.2011.169540

Fifth-generation digital immunoassay for prostate-specific antigen by single molecule array technology. / Wilson, David H.; Hanlon, David W.; Provuncher, Gail K.; Chang, Lei; Song, Linan; Patel, Purvish P.; Ferrell, Evan P.; Lepor, Herbert; Partin, Alan Wayne; Chan, Daniel Wan-Yui; Sokoll, Lori J; Cheli, Carol D.; Thiel, Robert P.; Fournier, David R.; Duffy, David C.

In: Clinical Chemistry, Vol. 57, No. 12, 12.2011, p. 1712-1721.

Research output: Contribution to journalArticle

Wilson, DH, Hanlon, DW, Provuncher, GK, Chang, L, Song, L, Patel, PP, Ferrell, EP, Lepor, H, Partin, AW, Chan, DW-Y, Sokoll, LJ, Cheli, CD, Thiel, RP, Fournier, DR & Duffy, DC 2011, 'Fifth-generation digital immunoassay for prostate-specific antigen by single molecule array technology', Clinical Chemistry, vol. 57, no. 12, pp. 1712-1721. https://doi.org/10.1373/clinchem.2011.169540
Wilson, David H. ; Hanlon, David W. ; Provuncher, Gail K. ; Chang, Lei ; Song, Linan ; Patel, Purvish P. ; Ferrell, Evan P. ; Lepor, Herbert ; Partin, Alan Wayne ; Chan, Daniel Wan-Yui ; Sokoll, Lori J ; Cheli, Carol D. ; Thiel, Robert P. ; Fournier, David R. ; Duffy, David C. / Fifth-generation digital immunoassay for prostate-specific antigen by single molecule array technology. In: Clinical Chemistry. 2011 ; Vol. 57, No. 12. pp. 1712-1721.
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