활성화된 Sepharose Gels에 공유결합으로 고정화된 Urokinase를 이용한 융합단백질 절단반응
- Title
- 활성화된 Sepharose Gels에 공유결합으로 고정화된 Urokinase를 이용한 융합단백질 절단반응
- Other Titles
- Fusion Protein Cleavage by Urokinase Covalentley Immobilized to Activated Sepharose Gels
- Author
- 이은규
- Keywords
- urokinase; immobilized enzyme; covalent bond; cleavage; fusion protein
- Issue Date
- 2000-03
- Publisher
- 한국생물공학회
- Citation
- 한국생물공학회지(KOREAN JOURNAL OF BIOTECHNOLOGY AND BIOENGINEERING), v. 15, no. 1, page. 42-48
- Abstract
- Urokinase (UK), a thrombolytic enzyme used to clear catheters obstructed by blood clots, can be also used industrially in
the recombinant protein purification system to cleave a fusion protein linked with a certain fragment of GST. We have
immobilized UK by covalent attachment to activated Sepharose 6B-Cl gels and evaluated its performance to cleave a fusion
protein of hGH and GST. The Sepharose gels were activated by etherification with glycidol (2,3-epoxypropanol) and further
oxidized with periodate resulting in glyceryl-Sepharose gels. After the activation treatment, surface density of the aldehyde
groups was 7-30 μmol-aldehyde/mL-gel. Immobilization yield was higher than 99% at high pH (10.5), and the immobilized
UK maintained ca. 80% specific activity of the soluble UK. In a column reaction the cleavage yield heavily depended on the
feed rate, and it was nearly 86% of that from soluble UK. And the immobilized UK was successfully regenerated by
unfolding and refolding with 6M GuHCl. After cleavaging reaction, the monomeric hGH was purified by using expanded bed
adsorption chromatography.
- URI
- https://www.dbpia.co.kr/journal/articleDetail?nodeId=NODE02246732https://repository.hanyang.ac.kr/handle/20.500.11754/161814
- ISSN
- 1225-7117; 2288-8268
- Appears in Collections:
- COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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