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Purification method for human plasma kallikrein by a new affinity chromatography
Biological & Pharmaceutical Bulletin
|March 26, 1998
Summary
Researchers developed PKSI-Toyopearl, a new affinity gel for rapidly purifying plasma kallikrein (PK). This method achieved a 1720-fold purification with an 85% yield, offering a valuable tool for plasma kallikrein isolation.
Area of Science:
- Biochemistry
- Protein Purification
- Biotechnology
Background:
- Plasma kallikrein is a key enzyme in the kallikrein-kinin system.
- Efficient purification methods are crucial for studying plasma kallikrein's function and therapeutic potential.
Purpose of the Study:
- To develop a novel affinity gel for the rapid and efficient purification of plasma kallikrein.
- To evaluate the efficacy of the synthesized PKSI-Toyopearl affinity gel.
Main Methods:
- Synthesis of PKSI-Toyopearl affinity gel using a selective plasma kallikrein inhibitor (PKSI-527).
- Purification of plasma kallikrein from kaolin-activated human plasma using the affinity gel.
- Further purification via DEAE-Toyopearl 650M chromatography.
- Analysis of purified protein using SDS-PAGE.
Main Results:
- The PKSI-Toyopearl gel enabled a rapid 181-fold purification of plasma kallikrein with an 85% yield.
- Subsequent chromatography resulted in a final purification of 1720-fold with a 63% yield.
- SDS-PAGE confirmed the presence of a single band at approximately 88 kDa.
Conclusions:
- PKSI-Toyopearl is an effective affinity ligand for the isolation of plasma kallikrein.
- This method provides a significant improvement in purification efficiency and speed.
- The developed technique is valuable for obtaining highly purified plasma kallikrein for research applications.