Precision Assay for Detection and Quantification of Anti-Prostate Specific Antigen Antibodies Using Indirect ELISA

Prostate-specific antigen (PSA) or gamma-selenoprotein or kallikrein-3 (KLK3) is a glycoprotein enzyme secreted by the epithelial cells of the prostate glands. It plays a crucial role in male fertility and is commonly used as a marker of prostate cancer. Antibodies to PSA antigen might play a role in male immune infertility. To date, the tests available in the market provide information only about the presence or absence of these antibodies in body fluids, which is further confirmed by the Western blot test. There are no tests available to quantify the amount of anti-PSA antibodies in human body fluid. Hence, the present patent relates to in vitro immunoassay for detecting and quantifying anti-prostate specific antigen (anti-PSA) antibodies in a human body fluid sample. In particular, the immunoassay is an indirect ELISA. The assay has demonstrated high sensitivity and specificity, capable of detecting anti-PSA antibodies in body fluids within a range of 4.61 ng/mL to 431.37 ng/mL. To further validate the assay's specificity, additional experiments have been conducted using various samples, including chicken seminal fluid, and the serum and semen of azoospermic individuals. These samples, standardized to the same volume, have been incubated with a fixed amount of human PSA-coated antigen. Similar experiments have been performed with anti-human, anti-rabbit, anti-mouse, anti-horse, and antigoat antibodies, further confirming the assay's specificity.