Research Product

This report describes development of an antibody to an organochlorine pesticide to be used in studies related to its inhibition of the ATPase system. Kelevan, the condensation product of ethyl levulinate and Kepone, was successfully conjugated to bovine serum albumin (BSA), fibrinogen (BF), and gamma globulin (BGG). Rabbits and chickens preimmunized with BSA and then immunized with BSA-Kelevan produced antibodies to both the hapten, Kelevan, and the carrier protein BSA. Antiserum to Kelevan protected ATPase activity against Kepone and its derivatives. The titer of antibody to Kelevan was critical since antiserum with only trace amounts of Kelevan antibody failed to protect the ATPase activity against Kepone inhibition. Antibody was concentrated by Na2SO4 fractional precipitation of the antiserum and obtained in pure form by affinity chromatography using BGG-Kel covalently linked to Sepharose 4B. Pure antibody was obtained from untreated blood serum or plasma with no prior pretreatment or fractionation using the BGG-Kel affinity column. Complete protection of mitochondrial Mg2 plus ATPase activity from in vitro inhibition of Kepone was obtained using a 1.2 mg quantity of Na2SO4 fractionated antibody and only 120 ug of pure antibody. Reversal of ATPase inhibition was readily obtained by addition of antibody prior to addition of substrate to the reaction mixture.