GENERATION OF ANTIBODIES AGAINST A HUMAN LIPOPROTEIN-LIPASE FUSION PROTEIN

Antibodies generated against specific proteins are useful tools for st udying the physiology and cell biology of the protein of interest. Alt hough antibodies have been successfully generated against lipoprotein lipase (LPL) and used to elucidate many aspects of its biology, there have been problems with the specificity, affinity and availability of these antibodies. To circumvent these problems, we have expressed a po rtion of human LPL as a bacterial fusion protein. The human LPL bacter ial fusion protein was utilized to generate polyclonal antibodies in r abbits that recognize intact human, rat and bovine LPL. Using these an tibodies, it was possible to demonstrate a direct correlation between LPL mass and LPL activity from different samples of human post-heparin plasma. In addition, these antibodies were used to develop an ELISA f or the measurement of LPL in tissue or plasma. This is a useful means for obtaining polyclonal antibodies to LPL in sufficient quantity and without contaminating mammalian proteins.