Isolation of urokinase by affinity ultrafiltration

Area of Science:

• Biochemistry
• Polymer Science
• Biotechnology

Background:

• Urokinase is a crucial plasminogen activator with significant therapeutic and diagnostic applications.
• Efficient and selective isolation methods for urokinase are essential for its purification and utilization.
• Existing purification techniques may be limited in yield or purity.

Purpose of the Study:

• To synthesize a novel water-soluble, ligand-bound polymer for urokinase isolation.
• To develop an affinity ultrafiltration process for purifying urokinase.
• To establish a new assay for urokinase activity using a chromogenic substrate.

Main Methods:

• Copolymerization of N-acryloyl-m-aminobenza-midicine and acrylamide to create an affinity polymer.
• Affinity ultrafiltration for isolating urokinase from artificial and crude urine solutions.
• Assay development by coupling plasminogen with L-benzoyl arginine-p-nitroanilide (L-BAPNA).

Main Results:

• Successful synthesis of a water-soluble, ligand-bound affinity polymer.
• Achieved urokinase isolation yields of 86% from artificial solutions and 49% from crude urine.
• Recovered urokinase demonstrated specific activity comparable to high-grade commercial standards.
• A new, cost-effective assay for urokinase was established.

Conclusions:

• The developed affinity polymer and ultrafiltration process provide an effective method for urokinase isolation.
• The purification strategy yields high-quality urokinase suitable for various applications.
• The new assay offers a valuable tool for quantifying urokinase activity.