Description
Description: This assay employs a two-site sandwich ELISA to quantitate EN2 in samples. An antibody specific for EN2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any EN2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for EN2 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of EN2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Prostate cancer (PC) is the second most common cause of cancer related death in men. A number of key limitations with prostate specific antigen (PSA), currently the standard detection test, has justified evaluation of new biomarkers.Engrailed-2 (EN2) protein, a homeodomain-containing transcription factor expressed in PC cell lines and secreted into the urine by PC in men. EN2 was expressed and secreted by PC cell lines and PC tissue but not by normal prostate tissue or stroma. The presence of EN2 in urine was highly predictive of PC, with a sensitivity of 66% and a specificity of 88.2%, without requirement for DRE. There was no correlation with PSA levels.Urinary EN2 is a highly specific and sensitive candidate biomarker of prostate cancer