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The continuous isolation of trypsin by affinity adsorbent recycling
A L Gill1, G W Niven, P G Scurlock
1Institute of Food Research, Reading Laboratory, Earley Gate, Whiteknights Road, Reading RG6 6BZ, UK; telephone: +118 9357214; fax: +118 9267917.
Biotechnology and Bioengineering
|July 22, 2008
Summary
This study introduces a continuous protein separation method using a polymer belt adsorbent. This novel approach enables rapid, diffusion-independent separation, achieving efficient trypsin isolation from complex mixtures.
Area of Science:
- Biotechnology
- Biochemical Engineering
- Protein Purification
Background:
- Continuous affinity separation methods are crucial for efficient protein purification.
- Existing methods often face limitations due to diffusion-dependent processes and slow cycle rates.
Purpose of the Study:
- To develop and evaluate a continuous affinity separation method utilizing a nonporous polymer belt adsorbent.
- To assess the efficiency of isolating trypsin from bovine pancreas using soybean trypsin inhibitor as the affinity ligand.
Main Methods:
- A polymer belt adsorbent was employed for continuous recycling through feedstock and eluent flows.
- Soybean trypsin inhibitor (SBTI) immobilized on nylon served as the affinity ligand for trypsin capture.
- The system was operated continuously for 30 hours to evaluate protein recovery and purity.
Main Results:
- Continuous isolation of trypsin from bovine pancreas crude preparation achieved 30-40% recovery over 30 hours.
- Approximately 18 mg of trypsin was obtained using a minimal amount of SBTI ligand.
- Chymotrypsin was identified as the primary contaminant, with purification rates dependent on contact time and protein concentration.
Conclusions:
- The nonporous belt adsorbent allows for rapid, diffusion-independent protein separation, enabling high cycle rates.
- Optimizing solid/liquid contact times and adsorption chamber design is key to maximizing purification rates and minimizing losses.
- This method demonstrates potential for efficient and scalable continuous protein purification.
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