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Z(basic) - A novel purification tag for efficient protein recovery
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0003-0140-419X
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0003-0605-8417
2007 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1161, no 1-2, 22-28 p.Article in journal (Refereed) Published
Abstract [en]

A positively charged protein domain, Z(basic) can be used as a general purification tag to achieve efficient recovery of recombinantly produced target proteins using cation-exchange chromatography. To construct a protein domain usable for ion-exchange chromatography, the surface of protein Z was engineered to be highly charged, which allowed for selective capture of target proteins on a cation-exchanger at physiological pH values. Interestingly, the novel domain, denoted Z(basic) was shown to be selective also under denaturing conditions and could preferably be used for purification of proteins solubilised from inclusion bodies. Moreover, a flexible process for solid-phase refolding was developed, using Z(basic) as a reversible linker to the cation-exchanger resin. This procedure has the inherited advantage of combining purification and refolding into a single step and still enabling elution of a concentrated product in a suitable buffer. This article summarizes development and use of the Z(basic), tag in small and pilot-plant-scale downstream processing.

Place, publisher, year, edition, pages
2007. Vol. 1161, no 1-2, 22-28 p.
Keyword [en]
Z(basic), ion-exchange chromatography, molecular modeling, proteolytic cleavage, purification of inclusion body proteins, solid-phase refolding
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-40724DOI: 10.1016/j.chroma.2007.05.091ISI: 000248891500005PubMedID: 17570380Scopus ID: 2-s2.0-34447617851OAI: oai:DiVA.org:kth-40724DiVA: diva2:442981
Note
26th International Symposium on the Separation of Proteins, Peptides and Polynucleotides Location: Innsbruck, Austria, Date: OCT 17-20, 2006Available from: 2011-09-23 Created: 2011-09-20 Last updated: 2017-12-08Bibliographically approved

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Hedhammar, MyHober, Sophia

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