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

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 a1-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%