D. Nadano et al., PURIFICATION OF MAMMALIAN RIBONUCLEASE USING IMMOBILIZED HUMAN RIBONUCLEASE INHIBITOR, Protein expression and purification, 7(2), 1996, pp. 167-172
Affinity chromatography on immobilized ribonuclease (RNase) inhibitor
was developed for purification of mammalian RNase. Human placental RNa
se inhibitor was conjugated to CNBr-activated Sepharose in the presenc
e of dithiothreitol. About 80% of the immobilized RNase inhibitor was
capable of binding bovine pancreatic RNase A. The bound RNase A was el
uted with 3 M NaCl at pH 5.0. Two 25-kDa and 1.8-kDa RNases, which wer
e obtained from human liver using a cellulose phosphate column, were b
ound to the immobilized RNase inhibitor and recovered in a pure and ac
tive form after treatment of the resin with p-hydroxymercuribenzoate.
These enzymes were considered to be nonsecretory-type RNases with diff
erent sugar contents. (C) 1996 Academic Press, Inc