Polymer–protein-enhanced fluoroimmunoassay for prostate-specific antigen

Abstract

Prostate-specific antigen (PSA) is a serum glycoprotein overproduced in prostate cancer, the total of which is comprised of two major forms, free and complexed. The common method for measuring of PSA is an Enzyme-linked immunosorbent assay (ELISA). Limits of detection using commercial PSA ELISA kits for free and total PSA were determined in our laboratory to be 1 and 10 ng/mL, respectively. A value of 0.10 was obtained for the free to total PSA ratio, a ratio of 0.25 being indicative of prostate cancer. A possible improvement in the sensitivity and detection limits of free and total PSA may be achieved by the ALYGNSA system (Clarizia et al., Anal Bioanal Chem 393:1531-1538, 2009). This fluorescent assay system employed a selective polymer biolinker system proven to enhance primary antibody orientation. Using this system, free and total PSA detection limits of 0.13 and 0.63 ng/mL, respectively, were realized. This amounted to an 8- and 15-fold improvement in detection limits for free and total PSA, respectively. A free to total PSA ratio of 0.20 was maintained in this study and may be useful for definitive diagnosis of prostate, as well as, other cancers.