Purification of an acidic recombinant protein from transgenic tobacco

✍ Scribed by Chris Holler; Chenming Zhang

• Publisher: John Wiley and Sons
• Year: 2008
• Tongue: English
• Weight: 234 KB
• Volume: 99
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.21638

✦ Synopsis

Abstract

Tobacco has proven to be a promising alternative for the production of recombinant therapeutic proteins and offers numerous advantages over other plants as a host system. However, the recovery and purification steps needed to obtain a protein at high recovery and purity have not been well investigated. In this study, a process was developed to purify a model acidic protein, recombinant β‐glucuronidase (rGUS) from transgenic tobacco leaf tissue, in three main steps after extraction: polyelectrolyte precipitation, hydrophobic interaction chromatography (HIC), and hydroxyapatite chromatography (HAC). Using this three‐step process, up to 40% of the initial rGUS activity could be recovered to near homogeneity as judged by SDS–PAGE. This work demonstrates that acidic recombinant proteins expressed in tobacco may be purified to high yield with high purity in a minimal amount of steps that are suitable for scale‐up. Furthermore, the general steps used in this process may suggest that a wide variety of acidic recombinant proteins may be purified in a similar manner from transgenic tobacco or other leafy crops. Biotechnol. Bioeng. 2008;99: 902–909. © 2007 Wiley Periodicals, Inc.