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Affinity chromatography of trypsin using chitosan as ligand support
1Laboratory of Enzyme Engineering, Agricultural College, Yangzhou University, China.
Journal of Chromatography. A
|August 23, 1996
Summary
Researchers developed a safer, simpler chitosan-based affinity adsorbent for protein purification. This new material effectively binds trypsin, showing promise for both lab and industrial-scale applications.
Area of Science:
- Biochemistry
- Materials Science
- Chemical Engineering
Background:
- Affinity chromatography is crucial for protein purification.
- Developing cost-effective and efficient adsorbent materials is an ongoing challenge.
- Chitosan offers a biocompatible and modifiable matrix for adsorbent development.
Purpose of the Study:
- To prepare and characterize chitosan-based affinity adsorbents for protein purification.
- To immobilize chicken ovomucoid onto chitosan beads for trypsin binding.
- To evaluate the performance and stability of the developed adsorbent.
Main Methods:
- Chitosan beads were synthesized and cross-linked with glutaraldehyde.
- Chicken ovomucoid was covalently immobilized onto the chitosan matrix.
- The binding capacity for trypsin was determined using enzyme activity assays.
- Chitosan-based columns were tested for trypsin chromatography and reusability.
Main Results:
- Successfully immobilized 60 mg of chicken ovomucoid per gram of chitosan.
- Achieved a maximum binding capacity of 8.10(4) U/g for trypsin.
- Demonstrated superior safety and simplicity compared to Sepharose-based methods.
- The affinity adsorbents exhibited good mechanical strength, stability, and reusability.
Conclusions:
- Chitosan is a suitable and advantageous matrix for developing affinity adsorbents.
- The developed chitosan-chicken ovomucoid adsorbent is effective for trypsin purification.
- This method offers a practical approach for both laboratory and large-scale protein purification.