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An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter
Published on: May 21, 2021
Separation and purification of enzymes by continuous pH-parametric pumping
S Y Huang1, C K Lin, L Y Juang
1Department of Chemical Engineering, National Taiwan University, Taipei, Taiwan, 107, Republic of China.
Biotechnology and Bioengineering
|October 1, 1985
Summary
Continuous pH-parametric pumping with affinity chromatography effectively purifies enzymes like trypsin and chymotrypsin. This method achieves high yields, demonstrating its efficiency for enzyme concentration and purification.
Area of Science:
- Biochemistry
- Chemical Engineering
- Chromatography
Background:
- Enzyme purification is crucial for various applications.
- Traditional methods can be time-consuming and less efficient.
- Developing continuous and high-yield purification techniques is essential.
Purpose of the Study:
- To develop and evaluate a continuous method for enzyme purification.
- To investigate the effectiveness of pH-parametric pumping coupled with affinity chromatography.
- To optimize conditions for concentrating and purifying trypsin, chymotrypsin, and stem bromelain.
Main Methods:
- Utilized continuous pH-parametric pumping for enzyme separation.
- Developed affinity adsorbents: chicken ovomucoid (CHOM) bound to chitin for trypsin, duck ovomucoid (DKOM) for chymotrypsin, and epsilon-Amino caproyl-D-tryptophan methyl ester coupled to chitosan for stem bromelain.
- Optimized pH levels for feed streams in the pumping process.
Main Results:
- Achieved high recovery rates, such as 95% for trypsin after 10 cycles.
- Demonstrated successful separation and purification of trypsin, chymotrypsin, and stem bromelain.
- Identified optimal operational conditions for the continuous purification process.
Conclusions:
- Continuous pH-parametric pumping combined with affinity chromatography is a highly effective technique for enzyme concentration and purification.
- This integrated approach offers high yields and efficiency compared to traditional methods.
- The study provides a robust platform for industrial-scale enzyme purification.
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