Development and validation of an ELISA to measure REG3E in canine blood

Regenerating islet-derived proteins (REG) are upregulated in people with sepsis, pancreatitis, and gastrointestinal diseases. One member of the REG family, namely REG3E, was recently identified in pancreatic tissue and plasma of dogs, with high expression in pancreatitis and sepsis. Our objective was to develop and validate an ELISA to measure REG3E concentrations in canine blood. An indirect sandwich ELISA was successfully developed using recombinant canine REG3E protein and polyclonal anti-canine REG3E antibodies raised in guinea pigs and rabbits. Antibody specificity was assessed using Western blot and mass spectrometric analysis of protein purified from canine plasma using Protein A Sepharose High Performance SpinTrap columns.