Monoclonal antibodies that distinguish between active and inactive forms of human postheparin plasma hepatic triglyceride lipase.
Hepatic triglyceride lipase (H-TGL) was purified from human postheparin plasma. Specific monoclonal antibodies (MAbs) were produced that discriminate between active (native) and inactive (denatured) forms of the enzyme. Mice immunized with native H-TGL resulted in MAbs that recognized only the native protein. The antibodies did not react with H-TGL treated with 1% sodium dodecyl sulfate or heated at 60 degrees C. The loss of immunoreactivity with heating correlated directly with the loss of enzyme activity and there was a corresponding increase in immunoreactivity with the MAbs prepared against the denatured enzyme. Western blot analysis of postheparin plasma with the MAbs against denatured H-TGL gave a single protein band of 65 kD; preheparin plasma showed no detectable immunoreactivity with either MAb. These immunochemical studies suggest that there are no circulating active or inactive forms of H-TGL in man. Furthermore, the MAbs provide the necessary reagents for development of immunoassays for H-TGL.
Mao
S J
SJ
Merrell Dow Research Institute, Cincinnati, OH 45215.
Rechtin
A E
AE
Jackson
R L
RL
D000911:Antibodies, Monoclonal; D006801:Humans; D008049:Lipase / Q000097:blood*; D008099:Liver / Q000201:enzymology*; D011233:Precipitin Tests; D013439:Sulfhydryl Reagents
D000911:Antibodies, Monoclonal; D013439:Sulfhydryl Reagents; D008049:Lipase