FELINE PANCREATIC LIPASE: PURIFICATION AND VALIDATION OF A CLINICALLY SIGNIFICANT RADIOIMMUNOASSAY FOR THE DIAGNOSIS OF FELINE PANCREATITIS A Thesis by BENJAMIN

Feline Pancreatic Lipase: Purification and Validation of a Clinically Significant Radioimmunoassay for the Diagnosis of Feline Pancreatitis. (December 2003) Benjamin Gregg Wilson, B.S., Texas A&M University Chair of Advisory Committee: Dr. Jörg M. Steiner Serum lipase activity has traditionally been used for diagnosis of pancreatitis in human beings and dogs. However, serum lipase activity is not specific for exocrine pancreatic function and many cell types other than pancreatic acinar cells also synthesize lipases. Recently, an immunoassay for the measurement of canine pancreatic lipase immunoreactivity has been developed and validated. This assay has shown to be specific for exocrine pancreatic function and sensitive for the diagnosis of canine pancreatitis. The objectives of this project were to purify feline pancreatic lipase (fPL), have antibodies against fPL (anti-fPL antibodies) produced, and develop a radioimmunoassay (RIA) for the diagnosis of feline pancreatitis. Pancreatic lipase was purified from feline pancreatic tissue by delipidation, anion-exchange chromatography, size-exclusion chromatography, and cation-exchange chromatography. Antiserum against fPL was raised in rabbits. Tracer was produced by iodination (I) of fPL using the chloramine T method. An RIA was established and validated by determination of sensitivity, dilutional parallelism, spiking recovery, intra-