Isolation and Characterization of alpha-1-Antitrypsin from Rhesus-Monkey Serum.

摘要

A simple, relatively gentle, procedure for isolation of rhesus-monkey alpah-1-antitrypsin from serum is described. The method consists of chromatographic separation of the fraction precipitated by 50-75%-satd. (NH4)2SO4 from pooled monkey serum on DEAE-cellulose followed by affinity chromatography on Sepharose-bound concanavalin A. Approx. 30% of the trypsin-inhibitory activity present in the original serum was recovered when alpha-1-antitrypsin was reconstituted with physiological saline (0.85% NaC1). Pure alpha-1-antitrypsin exhibited a single band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, with an estimated mol.wt. of 60000, and four bands in acid/starch-gel electrophoresis. The most rapidly migrating bands resembled the pattern and mobility for the normal human phenotype PiM in 28 Monkeys.