Development of a dual monoclonal antibody immunoassay for total human kallikrein 2

J. A. Finlay, J. R. Day, C. L. Evans, R. Carlson, K. Kuus-Reichel, L. S. Millar, S. D. Mikolajczyk, M. Goodmanson, G. G. Klee, H. G. Rittenhouse

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19 Scopus citations

Abstract

Background: Human kallikrein 2 (hK2) shares 80% sequence identity with prostate-specific antigen (PSA). Because both hK2 and hK2-α1-antichymotrypsin (hK2-ACT) complexes have been identified in patient sera, we devised an immunoassay for total hK2 [(thK2); hK2 and hK2-ACT] and evaluated it in healthy subjects and patients with prostate disease. Methods: We developed monoclonal antibodies (mAbs) with high specificity for hK2 and hK2-ACT and minimal cross-reactivity to PSA. Using these mAbs, a sandwich assay was developed and its specificity for forms of hK2 was assessed. Serum samples (n = 1035) from healthy volunteers, patients with increased PSA, and men who had undergone radical prostatectomy were assayed for thK2. We also measured thK2 in samples before and after storage under common laboratory conditions. Results: The minimum detectable concentration in the thK2 assay was 0.008μg/L, and PSA cross-reactivity was <0.001%. The assay detected prohK2 and three different hK2-serum protease complexes. The median serum concentration of thK2 in control samples (0.013 μg/L) was significantly lower than the median in samples from patients with increased PSA concentrations (0.085 μg/L). Immunoreactive hK2 changed little in samples stored for up to 1 month at -70°C. Conclusions: The thK2 assay recognizes all forms of hK2 that have been found in bodily fluids to date.

Original languageEnglish (US)
Pages (from-to)1218-1224
Number of pages7
JournalClinical chemistry
Volume47
Issue number7
DOIs
StatePublished - 2001

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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    Finlay, J. A., Day, J. R., Evans, C. L., Carlson, R., Kuus-Reichel, K., Millar, L. S., Mikolajczyk, S. D., Goodmanson, M., Klee, G. G., & Rittenhouse, H. G. (2001). Development of a dual monoclonal antibody immunoassay for total human kallikrein 2. Clinical chemistry, 47(7), 1218-1224. https://doi.org/10.1093/clinchem/47.7.1218