Large scale, rapid purification of recombinant tissue‐type plasminogen activator

FEBS Letters

Newsome, P.¹  Browne, M.J.¹  Dodd, I.¹  Robinson, J.H.¹  Southwick, W.¹  Jalalpour, S.¹ 

[1] Beecham Pharmaceuticals Research Division, Biosciences Research Centre, Great Burgh, Yew Tree Bottom Road, Epsom, Surrey KT18 5XQ, England

关键词: Recombinant t-PA Zinc chelate-Sepharose CL4B Lysine-Sepharose CL4B Continuous chromatography;   

DOI  :  10.1016/0014-5793(86)81075-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Recombinant tissue-type plasminogen activator (rt-PA) from cultures of a genetically manipulated Bowes melanoma cell line (TRBM6) was purified in batches of average volume 451 using an autoclavable, reusable, continuous chromatography system comprising zinc chelate-Sepharose CL4B and lysine-Sepharose CL4B. After eight successive purifications the rt-PA was ultrafiltered to yield a preparation containing 4.9 mg protein/ml and 2.7 × 10⁶ . Analysis by SDS-polyacrylamide gel electrophoresis followed by staining with Coomassie brilliant blue R250 showed major protein bands at M _r = 63000 and 65000; most of the material was in the 1-chain form. The potential usefulness of a simple, rapid continuous chromatography system that can be operated under aseptic conditions is discussed.

【 授权许可】

Unknown