Premium
Isolation of urokinase by affinity ultrafiltration
Author(s) -
Male K. B.,
Nguyen A. L.,
Luong J. H. T.
Publication year - 1990
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260350112
Subject(s) - urokinase , chromogenic , chemistry , ultrafiltration (renal) , substrate (aquarium) , acrylamide , chromatography , polymer , polymer chemistry , nuclear chemistry , monomer , organic chemistry , biology , surgery , medicine , ecology
A water‐soluble, ligand‐bound polymer has been synthesized for the purpose of isolation of urokinase, an important plasminogen activator. The affinity polymer was formed by copolymerizing N ‐acryloyl‐ m ‐aminobenza‐midine and acrylamide in the absence of oxygen. An affinity ultrafiltration process was then developed for isolating urokinase from an artificial solution containing peroxidase and urokinase and from a crude urine source. The process yields were determined to be 86% and 49%, respectively. The recovered urokinase exhibited a specfic activity close to that of the highest commercial grade. This article also presents a new technique for assaying urokinase by coupling plasminogen with L‐benzoyl arginine‐ p ‐nitroanilide (L‐BAPNA), an inexpensive chromogenic substrate.