Large scale, rapid purification of recombinant tissue-type plasminogen activator.

Abstract

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 X 10(6) IU/ml. Analysis by SDS-polyacrylamide gel electrophoresis followed by staining with Coomassie brilliant blue R250 showed major protein bands at Mr = 63,000 and 65,000; 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.

• Authors: I Dodd, S Jalalpour, W Southwick, P Newsome, M J Browne, J H Robinson
• Journal: FEBS letters (FEBS Lett) Vol. 209 Issue 1 Pg. 13-7 (Dec 01 1986) ISSN: 0014-5793 [Print] England
• PMID: 3100325 (Publication Type: Journal Article)

Chemical References

• Recombinant Proteins
• Tissue Plasminogen Activator

Topics

• Animals
• Cell Line
• Chromatography, Gel
• Electrophoresis, Polyacrylamide Gel
• Melanoma, Experimental
• Molecular Weight
• Recombinant Proteins (isolation & purification)
• Tissue Plasminogen Activator (isolation & purification)