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[Isolation and purification of urokinase]
Voprosy Meditsinskoi Khimii
|September 1, 1981
Summary
Researchers developed a new method to isolate urokinase, a key enzyme, from human urine using ion exchange chromatography. This improved procedure yields a highly pure urokinase preparation for potential therapeutic applications.
Area of Science:
- Biochemistry
- Protein purification
- Enzymology
Context:
- Urokinase is a vital enzyme found in human urine.
- Existing methods for urokinase isolation can be inefficient or yield impure products.
- The need for a reliable and scalable method for obtaining pure urokinase is critical for research and therapeutic use.
Purpose:
- To develop an improved and efficient procedure for the isolation of urokinase from human urine.
- To achieve a chromatographically pure urokinase preparation.
- To optimize the ion exchange chromatography and depigmentation steps for urokinase purification.
Summary:
- An enhanced protocol for urokinase isolation from human urine was established utilizing ion exchange chromatography on carboxyl cationite SMT at pH 4.7.
- Depigmentation of urine was effectively performed using anionite FAF pretreatment.
- Subsequent stepwise elution with a phosphate buffer at pH 7.0, followed by gel filtration through Sephadex G-100 and G-150, resulted in a chromatographically pure urokinase preparation.
Impact:
- This improved method provides a more efficient way to obtain high-purity urokinase.
- The purified urokinase can be utilized in various biomedical research applications.
- The protocol's success in achieving high purity suggests potential for therapeutic applications of the isolated urokinase.