Abstract

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. (c) 2007 Elsevier B.V. All rights reserved

Published in

Journal of Chromatography A
2007, Volume: 1161, number: 1-2, pages: 22-28
Publisher: ELSEVIER SCIENCE BV

SLU Authors

UKÄ Subject classification

Animal and Dairy Science
Veterinary Science

Publication Identifiers

DOI: https://doi.org/10.1016/j.chroma.2007.05.091

Permanent link to this page (URI)

https://res.slu.se/id/publ/16747