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Purification of an acidic recombinant protein from transgenic tobacco
Author(s) -
Holler Chris,
Zhang Chenming
Publication year - 2008
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21638
Subject(s) - recombinant dna , protein purification , transgene , biochemistry , affinity chromatography , chemistry , chromatography , tobacco etch virus , biology , enzyme , gene , plant virus , virus , virology , potyvirus
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.