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Zbasic: a purification tag for selective ion-exchange recovery
KTH, School of Biotechnology (BIO), Protein Technology.ORCID iD: 0000-0003-0140-419X
KTH, School of Biotechnology (BIO), Protein Technology.ORCID iD: 0000-0002-6104-6446
KTH, School of Biotechnology (BIO), Protein Technology.ORCID iD: 0000-0003-0605-8417
2014 (English)In: Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029, Vol. 1129, 197-204 p.Article in journal (Refereed) Published
Abstract [en]

A positively charged protein domain, denoted Zbasic, can be used as a general purification tag for purification of recombinantly produced target proteins by cation-exchange chromatography. The Zbasic domain is constructed from the Protein A-derived domain and engineered to be highly charged, which allows selective capture on a cation exchanger at physiological pH values. Moreover, Zbasic is selective also under denaturing conditions and can be used for purification of proteins solubilized from inclusion bodies. Zbasic can then be used as a flexible linker to the cation-exchange resin and thereby allow solid-phase refolding of the target protein.Herein, protocols for purification of soluble Zbasic-tagged fusion proteins, as well as for integrated purification and solid-phase refolding of insoluble fusion proteins, are described. In addition, a procedure for enzymatic tag removal and recovery of native target protein is outlined.

Place, publisher, year, edition, pages
2014. Vol. 1129, 197-204 p.
National Category
Biochemistry and Molecular Biology
Research subject
URN: urn:nbn:se:kth:diva-182468OAI: diva2:904566

QC 20160303

Available from: 2016-02-19 Created: 2016-02-19 Last updated: 2016-03-03Bibliographically approved

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Hedhammar, MyNilvebrant, JohanHober, Sophia
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