Lipopeptide adjuvants: Generation of lactate dehydrogenase isoenzyme-specific antibodies for immunochemical diagnosis

Lactate dehydrogenase catalyzes the final step in glycolysis, th interconve rsion of pyruvate and lactate. The tetrameric enzyme is composed of one or two subunits (H and/or M) resulting in five isoenzyme forms: LDH-H4, -H3M1, -H2M2, -H1M3, and -M4. The relative distribution of the LDH isoenzymes is tissue dependent and a significant marker for the diagnosis of hepatoma of the liver, myocardial infarction, muscular dystrophy, and a wide variety of other acute and chronic diseases to be detected by alterations of the LDH isoenzyme pattern in serum. Immunochemical approaches to the routine determ ination of LDH depend on isoenzyme specific antibodies. Since the H- and M- subunits for human LDH are highly homologous, LDH isoenzyme specific antibo dies for immunochemical monitoring are hard to generate. Here we present da ta on the generation and characterization of LDH isoenzyme-specific mono- a nd polyclonal antibodies in different species in the presence of lipopeptid e adjuvants. Western-Blot and ELISA analysis showed that antisera and monoc lonal antibodies recognize their homologous antigens with high specificity and are therefore suitable for immunochemical monitoring of the LDH isoenzy mes H4 an M4. In addition, they can be used for the determination of LDH is oenzyme specific activity which is an essential prerequisite for online amp erometric immunosensor monitoring.