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Development and Analytical Validation of a Radioimmunoassay for the Measurement of Trypsin-like Immunoreactivity in Ferret Serum
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Ferrets are valuable animal models for the study of many human diseases. Cystic fibrosis (CF) and pancreatitis are two such human diseases that could benefit from a ferret model. The pancreatic disease processes that occur in the ferret animal model may allow researchers to investigate the pathophysiology of these diseases and help to develop new therapeutics for them. Trypsin-like immunoreactivity (TLI) is a noninvasive diagnostic tool used to assess exocrine pancreatic function in humans, dogs, and cats. Therefore, the major objective of this project was to purify trypsin from ferret pancreas and to set up and analytically validate a radioimmunoassay (RIA) for the measurement of TLI in ferret serum. Ferrets euthanized for unrelated projects had their pancreata removed and frozen at -80⁰C for further purification. Ferret trypsin was purified from pancreatic tissue and antiserum against ferret trypsin was raised in rabbits. Tracer was produced by the chloramine-T method. A radioimmunoassay was set up and analytically validated by determination of dilutional parallelism, spiking recovery, intra-assay variability, interassay variability, and sensitivity. A reference interval was established from 35 healthy ferret serum samples. A small group of ferrets with acute pancreatitis (AP; n = 3) and healthy controls (HC; n = 4) were analyzed by the new TLI RIA. We conclude that the ferret TLI RIA is analytically sensitive, accurate, precise, reproducible, and sufficiently linear for the measurement of TLI in serum samples from ferrets. All AP ferrets had higher serum TLI concentrations than HC ferrets, although the sample size was small. Further studies evaluating the usefulness of measuring serum ferret TLI concentrations in animal models with AP and CF are underway.
Bridges, Cory Seth (2017). Development and Analytical Validation of a Radioimmunoassay for the Measurement of Trypsin-like Immunoreactivity in Ferret Serum. Master's thesis, Texas A & M University. Available electronically from