ENSYME IMMUNOASSAY FOR 8-ISO-2,3-DINOR PROSTAGLADIN F1

The long-term objective of this proposal is the development of a reliable and sensitive assay for the accurate measurement of lipid peroxidation as an index of oxidative stress. The implication of oxidative stress in atherosclerosis, ischemia-reperfusion injury, inflammation, cancer, and aging highlights the importance of establishing methodology for the measurement of a reliable market or peroxidative injury. Recent identification of 8-iso-2,3-dinor PGF1alpha as a reliable marker of lipid peroxidation makes it an ideal target for the development of an immunoassay. Specific antibodies that recognize 8-iso-2,3-dinor PGF1alpha are vital for the development of an immunoassay. Antibody development is the primary objective of the present proposal. This will be achieved by a) the synthesis of large quantities of 8-iso-2,3-dinor PGF1alpha and transforming it into a structural analog, b) conjugating 8-iso-2,3-dinor PGF1alpha and its structural analog to a carrier protein, c) injecting in to rabbits and mice, and d) screening the antisera for specific antibodies using an acetylcholinesterase conjugate of 8-iso-2,3-dinor PGF1alpha. Discernable titers of the antibody for 8-iso-2,3-dinor PGF1alpha are expected towards the end of the Phase I contract period. Any highly specific polyclonal antibodies obtained from rabbits will be used for the development of an enzyme immunoassay. Mice producing high titer antisera will be used to develop monoclonal antibodies.