TY - JOUR AU - 이은규 DA - 2000/03 PY - 2000 UR - https://www.dbpia.co.kr/journal/articleDetail?nodeId=NODE02246732 UR - https://repository.hanyang.ac.kr/handle/20.500.11754/161814 AB - 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. PB - 한국생물공학회 KW - urokinase KW - immobilized enzyme KW - covalent bond KW - cleavage KW - fusion protein TI - 활성화된 Sepharose Gels에 공유결합으로 고정화된 Urokinase를 이용한 융합단백질 절단반응 TT - Fusion Protein Cleavage by Urokinase Covalentley Immobilized to Activated Sepharose Gels T2 - 한국생물공학회지(KOREAN JOURNAL OF BIOTECHNOLOGY AND BIOENGINEERING) ER -