Enzymatic cleavage of fusion proteins using immobilised protease 3C
2006 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 47, no 2, 422-426 p.Article in journal (Refereed) Published
A strategy for efficient cleavage of fusion proteins using an immobilised protease has been developed. Protease 3C from coxsackie virus was recombinantly produced in Escherichia coli and covalently immobilised onto a solid support. Thereafter, Z(basic) tagged fusion proteins, with a specific cleavage sequence between the domains, were flown through the proteolytic column and circulated until complete cleavage. Subsequently, the processed protein solution was applied on a cation exchanger. Thereby, removal of the released, positively charged fusion tag, Z(basic), was done by adsorption to the matrix while the target proteins were recovered in the flow through. Interestingly, the columns were shown to be reusable without any measurable decrease in activity. Moreover, after storage in 4 degrees C for two months the activity was almost unaffected.
Place, publisher, year, edition, pages
2006. Vol. 47, no 2, 422-426 p.
immobilised protease, Z(basic), protease 3C, cleavage, purification, efficient
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-15745DOI: 10.1016/j.pep.2006.01.003ISI: 000238277000011ScopusID: 2-s2.0-33646541425OAI: oai:DiVA.org:kth-15745DiVA: diva2:333787
QC 201005252010-08-052010-08-052010-10-20Bibliographically approved