Title:
コラゲナーゼを精製する方法
Document Type and Number:
Japanese Patent JP3662020
Kind Code:
B2
Abstract:
A process for purifying crude collagenase is disclosed. The collagenase purification process includes providing a stabilized crude collagenase solution containing collagenase, pigment, toxins, bacterial materials, and proteolytic enzyme impurities including clostripain, trypsin, and caseinase. The stabilized collagenase solution is applied to hydroxylapatite packing. pigment and caseinase are eluted with a first solution comprising about 0.05 M to about 0.3 M phosphate buffer, and then collagenase, trypsin, and clostripain are eluted with a second solution comprising about 0.35 M to about 0.5 M phosphate buffer to provide a first collected solution. The first collected solution is then applied to gel filtration packing and collagenase and clostripain are eluted with a neutral pH buffer solution, to provide a second collected solution. The second collected solution is then applied to Reactive Red 120-Agarose packing and collagenase is eluted with a neutral pH buffer solution to provide purified collagenase. The process provides extremely pure collagenase in high yield with reduced consumption of eluting solutions and avoids unpredictable gradient eluting techniques.
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Inventors:
Lee, Catherine
Honasek, Cynthia
Din, Tan, Sun
Honasek, Cynthia
Din, Tan, Sun
Application Number:
JP52343294A
Publication Date:
June 22, 2005
Filing Date:
April 13, 1994
Export Citation:
Assignee:
Edwards Lifesciences Corporation
International Classes:
C07K1/16; C12N9/52; C12R1/145; (IPC1-7): C12N9/52
Other References:
Biochemistry, 1984年, Vol.23, p.3077-3085
Attorney, Agent or Firm:
Hidesaku Yamamoto
Takaaki Yasumura
Natsuki Morishita
Takaaki Yasumura
Natsuki Morishita