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Highly Sensitive Automated Chemiluminometric Assay for Measuring Free Human Glandular Kallikrein-2

Departments of
¹ Laboratory Medicine and Pathology,
² Epidemiology, and
³ Urology Research, Mayo Clinic and Mayo Foundation, 200 First Street SW, Rochester, MN 55905.
⁴ Department of Research and Development, Hybritech Inc., a subsidiary of Beckman Coulter, Inc., San Diego, CA 92121.
^a Author for correspondence. Fax 507-284-4542; e-mail klee.george{at}mayo.edu

Background: Human glandular kallikrein (hK2) is a serine protease that has 79% amino acid identity with prostate-specific antigen (PSA). Both free hK2 and hK2 complexed to ₁-antichymotrypsin (ACT) are present in the blood in low concentrations. We wished to measure hK2 in serum with limited contribution from hK2-ACT for the results.

Methods: We developed an automated assay for hK2 with use of a select pair of monoclonal antibodies. The prototype assay was implemented on a Beckman Coulter ACCESS^® analyzer.

Results: The detection limit of the assay was 1.5 ng/L, the "functional sensitivity" (day-to-day CV <15%) was <4 ng/L, cross-reactivity with PSA and PSA-ACT was negligible, and cross-reactivity with hK2-ACT was 2%. After surgical removal of prostate glands, serum hK2 was <7 ng/L and was <15 ng/L in most healthy women. The median serum concentration of hK2 in healthy men without prostate cancer was 26 ng/L. The median concentration of hK2 was 72 ng/L for men having prostate cancer with lower Gleason scores compared with 116 ng/L for men with more advanced cancer. The concentration of hK2 correlated weakly with PSA, with the mean hK2 concentrations generally 30- to 80-fold lower than PSA concentrations.

Conclusion: The availability of a robust, high sensitivity automated assay for hK2 should facilitate further investigations of the role of hK2 measurements in the management of patients with prostate disease.© 1999 American Association for Clinical Chemistry

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