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Development of Sensitive Immunoassays for Free and Total Human Glandular Kallikrein 2

¹ University of Turku, Department of Biotechnology, Turku, Finland.
² Memorial Sloan-Kettering Cancer Center, New York, NY.
³ Lund University, Department of Laboratory Medicine, Division of Clinical Chemistry, University Hospital UMAS, Malmö, Sweden.
⁴ University Hospital of Turku, Department of Surgery, Turku, Finland.

^aAddress correspondence to this author at: University of Turku, Department of Biotechnology, Tykistökatu 6 A 6th Floor, 20520 Turku, Finland. Fax 358-2-333-8050; e-mail ville.vaisanen{at}pp.nic.fi.

Background: Free and total human kallikrein 2 (hK2) might improve the discrimination between prostate cancer and benign prostatic hyperplasia. Concentrations of hK2 are 100-fold lower than concentrations of prostate-specific antigen (PSA); therefore, an hK2 assay must have a low detection limit and good specificity.

Methods: PSA- and hK2-specific monoclonal antibodies were used in solid-phase, two-site immunofluorometric assays to detect free and total hK2. The total hK2 assay used PSA-specific antibodies to block nonspecific signal. The capture antibody of the free hK2 assay did not cross-react with PSA. To determine the hK2 concentrations in the male bloodstream, total hK2 was measured in a control group consisting of 426 noncharacterized serum samples. Free and total hK2 were measured in plasma from 103 patients with confirmed prostate cancer.

Results: All 426 males in the control group had a total hK2 concentration above the detection limit of 0.0008 µg/L. The median total hK2 concentration was 0.022 µg/L (range, 0.0015–0.37 µg/L). hK2 concentrations were 0.1–58% of total PSA (median, 3.6%). hK2 concentrations were similar in men 41–50 and 51–60 years of age. The ratio of hK2 to PSA steadily decreased from 5–30% at PSA <1 µg/L to 1–2% at higher PSA concentrations. In 103 patients with prostate cancer, the median hK2 concentration in plasma was 0.079 µg/L (range, 0.0015–16.2 µg/L). The median free hK2 concentration was 0.070 (range, 0.005–12.2) µg/L. The proportion of free to total hK2 varied from 17% to 131% (mean, 85%).

Conclusions: The wide variation in the free-to-total hK2 ratio suggests that hK2 in blood plasma is not consistently in the free, noncomplexed form in patients with prostate cancer. The new assay is sufficiently sensitive to be used to study the diagnostic accuracies of free and total hK2 for prostate cancer.

The following articles in journals at HighWire Press have cited this article:

				T. Steuber, A. J. Vickers, A. M. Serio, V. Vaisanen, A. Haese, K. Pettersson, J. A. Eastham, P. T. Scardino, H. Huland, and H. Lilja Comparison of Free and Total Forms of Serum Human Kallikrein 2 and Prostate-Specific Antigen for Prediction of Locally Advanced and Recurrent Prostate Cancer Clin. Chem., February 1, 2007; 53(2): 233 - 240. [Abstract] [Full Text] [PDF]