Purification and partial characterization of α1-proteinase inhibitor in the common marmoset (Callithrix jacchus)

Fecal alphal-proteinase inhibitor (alpha(1)-PI) concentration has been to diagnose enteric protein loss in dogs and cats. Chronic lymphocytic enteritis is commonly seen in the marmoset (Callithrix jaccus) and is characterized by hypoalbuminemia. As a prelude to immunoassay development for detecting enteric protein loss, marmoset serum alpha(1)-PI was purified using immunoaffinity chromatography and ceramic hydroxyapatite chromatography. Partial characterization was performed by reducing gel electrophoresis and enzyme inhibitory assays. Protein identity was confirmed with peptide mass fingerprinting and N-terminal amino acid sequencing. Molecular mass, relative molecular mass, and isoelectric point for marmoset alpha(1)-PI were 54 kDa, 51,677, and 4.8-5.4, respectively. Trypsin, chymotrypsin, and elastase inhibitory activity were observed. N-terminal amino acid sequence for marmoset alpha(1)-PI was EDPQGDAAQKMDTSHH. In conclusion, marmoset alpha(1)-PI was successfully purified from serum with an overall yield of 12% using a rapid and efficient method. Purified marmoset alpha(1)-PI has characteristics similar to those of alpha(1)-PI reported for other species. (C) 2015 Elsevier Ltd. All rights reserved.