A Monoclonal Antibody Specific for Δ12-Prostaglandin J2 and Its Utilization in the Immunological Assay in Cell Culture System of Adipocytes

Prostaglandin (PG) D2 can be produced in adipocytes and dehydrated to PGs of J2 series, including Δ12-PGJ2 and 15-deoxy-Δ12,14-PGJ2 (15d-PGJ2), which serve as pro-adipogenic prostanoids through the activation of peroxisome proliferator-activated receptor γ. To accomplish the quantification of Δ12-PGJ2 in the cell culture system of adipocytes, the present study aimed to develop a sensitive and specific immunological assay for Δ12-PGJ2. Here, we established a cloned hybridoma cell line secreting a monoclonal antibody specifically recognizing Δ12-PGJ2 and utilized for the development of its solid-phase enzyme-linked immunosorbent assay (ELISA). The immobilized antigen using a conjugate of Δ12-PGJ2 and γ-globulin was competitively allowed to react with the monoclonal antibody in the presence of free Δ12-PGJ2. The assay provided a sensitive calibration curve for Δ12-PGJ2, allowing us to determine a range from 0.16 pg to 0.99 ng with a value of 13 pg at 50% displacement in one assay. The monoclonal antibody sho...