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Clinical Chemistry 47: 1218-1224, 2001;
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(Clinical Chemistry. 2001;47:1218-1224.)
© 2001 American Association for Clinical Chemistry, Inc.

Articles

Development of a Dual Monoclonal Antibody Immunoassay for Total Human Kallikrein 2

Judith A. Finlay1a, John R. Day1, Cindy L. Evans1, Robert Carlson1, Kristine Kuus-Reichel1, Lisa S. Millar1, Stephen D. Mikolajczyk1, Marcia Goodmanson2, George G. Klee2 and Harry G. Rittenhouse1

1 Research Department, Hybritech Inc., a subsidiary of Beckman Coulter, Inc., San Diego, CA 92121.

2 Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN 55905.


aAddress correspondence to this author at: PO Box 269006, San Diego, CA 92196-9006. Fax 619-621-4610; e-mail jafinlay{at}beckman.com

Background: Human kallikrein 2 (hK2) shares 80% sequence identity with prostate-specific antigen (PSA). Because both hK2 and hK2-{alpha}1-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 0.008 µg/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.




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


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V. Vaisanen, S. Eriksson, K. K. Ivaska, H. Lilja, M. Nurmi, and K. Pettersson
Development of Sensitive Immunoassays for Free and Total Human Glandular Kallikrein 2
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S. D. Mikolajczyk, W. J. Catalona, C. L. Evans, H. J. Linton, L. S. Millar, K. M. Marker, D. Katir, A. Amirkhan, and H. G. Rittenhouse
Proenzyme Forms of Prostate-Specific Antigen in Serum Improve the Detection of Prostate Cancer
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R. K. Nam, W. W. Zhang, J. Trachtenberg, E. Diamandis, A. Toi, M. Emami, M. Ho, J. Sweet, A. Evans, M. A.S. Jewett, et al.
Single Nucleotide Polymorphism of the Human Kallikrein-2 Gene Highly Correlates With Serum Human Kallikrein-2 Levels and in Combination Enhances Prostate Cancer Detection
J. Clin. Oncol., June 15, 2003; 21(12): 2312 - 2319.
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A. Haese, V. Vaisanen, J. A. Finlay, K. Pettersson, H. G. Rittenhouse, A. W. Partin, D. J. Bruzek, L. J. Sokoll, H. Lilja, and D. W. Chan
Standardization of Two Immunoassays for Human Glandular Kallikrein 2
Clin. Chem., April 1, 2003; 49(4): 601 - 610.
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B. G. Blijenberg, M. F. Wildhagen, C. H. Bangma, J. A. Finlay, V. Vaisanen, and F. H. Schroder
Comparison of Two Assays for Human Kallikrein 2
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